BR112012013876B1 - Isolated antibody or a fragment thereof and pharmaceutical composition comprising the same - Google Patents

Abstract

isolated antibody or a fragment thereof and pharmaceutical composition comprising the present disclosure provides compositions and methods related to or derived from antigen binding proteins that activate fgf21-mediated signaling. in modalities, antigen binding proteins specifically bind to (i) ß-klotho; (ii) fgfr1c, fgfr2c, fgfr3c or fgfr4; or (iii) a complex comprising ß-klotho and one of fgfr1c, fgfr2c, fgfr3c and fgfr4. in some embodiments, antigen-binding proteins induce fgf21-like signaling. in some embodiments, antigen binding proteins are fully human, humanized or chimeric antibodies, fragments and derivatives binding those antibodies, and polypeptides that specifically bind to (i) ß-klotho; (ii) fgfr1c, fgfr2c, fgfr3c or fgfr4; or (iii) a complex comprising ß-klotho and one of fgfr1c, fgfr2c, fgfr3c and fgfr4. other modalities provide nucleic acids that encode these antigen binding proteins, and fragments and derivatives thereof, and polypeptides, cells that comprise these polynucleotides, methods of producing those antigen binding proteins, and fragments and derivatives thereof, and polypeptides, and methods of use of these antigen-binding proteins, fragments and derivatives thereof, and polypeptides, including methods of treatment or diagnosis of individuals suffering from type 2 diabetes, obesity, nash, metabolic syndrome and related disorders or conditions.

Description

ISOLATED ANTIBODY OR A FRAGMENT OF THE SAME AND PHARMACEUTICAL COMPOSITION THAT UNDERSTANDS THE SAME

This application claims the benefit of Provisional U.S. Application No. 61 / 267,321, filed on December 7, 2009, and Provisional U.S. Application No. 61 / 381,846, filed on September 10, 2,010, which are hereby incorporated by reference.

SEQUENCE LISTING

This application contains a Sequence Listing that was submitted in ASCII format through EFS-Web and is incorporated herein by reference in its entirety. Said ASCII copy, created on November 24, 2010, is called A-1519-WO-PCT.txt and is 645,894 bytes in size.

FIELD OF THE INVENTION

The present disclosure relates to the nucleic acid molecules that encode antigen binding proteins that bind to (i) β-Klotho; (ii) FGFR1c, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFR1c, FGFR2c, FGFR3c and FGFR4. The present disclosure also provides antigen binding proteins that bind to (i) β-Klotho; (ii) FGFR1c, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFR1c, FGFR2c, FGFR3c and FGFR4, including antigen binding proteins that induce FGF21-like (FGF21-like) signaling, in addition to pharmaceutical compositions comprising binding proteins antigens that bind to (i) β-Klotho; (ii) FGFR1c, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFR1c, FGFR2c, FGFR3c and FGFR4, including protein-binding proteins

Petition 870190023322, of 03/11/2019, p. 12/18 antigens that induce FGF21 type signaling, and methods for the treatment of metabolic disorders with the use of these nucleic acids, polypeptides or pharmaceutical compositions. Diagnostic methods using antigen-binding proteins are also provided.

BACKGROUND OF THE INVENTION fibroblast growth factor 21 (FGF21) is a secreted polypeptide that belongs to a subfamily of fibroblast growth factors (FGFs) that includes FGF19, FGF21 and FGF23 (Itoh et al., (2004) Trend Genet. 20 : 563-69). FGF21 is an atypical FGF in that it is independent of heparin and functions as a hormone in the regulation of glucose, lipid and energy metabolism.

It is highly expressed in the liver and pancreas and is the only member of the FGF family to be primarily expressed in the liver. Transgenic mice that overexpress FGF21 exhibit metabolic phenotypes of slow growth rate, low plasma glucose and triglyceride levels, and an absence of type 2 diabetes associated with age, islet hyperplasia and obesity. Pharmacological administration of recombinant FGF21 protein in rodent and primate models results in normalized blood glucose levels, reduced triglyceride and cholesterol levels and increased glucose tolerance and insulin sensitivity. In addition, FGF21 reduces body weight and body fat by increasing energy expenditure, physical activity and metabolic rate. Experimental research provides support for the pharmacological administration of FGF21 for the treatment of type 2 diabetes, obesity, dyslipidemia and other metabolic conditions or disorders in humans.

FGF21 is an endocrine hormone derived from the liver that stimulates glucose uptake in adipocytes and lipid homeostasis through activation of its receptor. Interestingly, in addition to the canonical FGF receptor, the FGF21 receptor also understands membrane-associated β-Klotho as an essential cofactor. Activation of the FGF21 receptor leads to multiple effects on several metabolic parameters.

In mammals, FGFs mediate their action through a set of four FGF receptors, FGFR1 - 4, which, in turn, are expressed in multiple splicing variants, for example, FGFRlc, FGFR2c, FGFR3c and FGFR4. Each FGF receptor contains an intracellular tyrosine kinase domain that is activated by ligand binding, leading to downstream signaling pathways involving MAPKs (Erkl / 2), RAFI, AKT1 and STATs (Kharitonenkov et al., (2008) BioDrugs 22 : 37-44). Several reports suggest that FGFR1-3 c-reporter splicing variants exhibit specific affinity for β-Klotho and could act as an endogenous receptor for FGF21 (Kurosu et al., (2007) J. Biol. Chem. 282: 26.687-26.695 ; Ogawa et al., (2007) Proc. Natl. Acad. Sci. USA 104: 7.432-7.437; Kharitonenkov et al., (2008) J. Cell. Physiol. 215: 1-7). In the liver, which abundantly expresses both β-Klotho and FGFR4, FGF21 does not induce MAPK phosphorylation, despite the strong binding of FGF21 to the p-Klotho-FGFR4 complex. In 3T3-L1 cells and white adipose tissue, FGFR1 is by far the most abundant receptor and it is therefore more likely that the main functional FGF21 receptors in this tissue are the βKlotho / FGFRlc complexes.

The present disclosure provides a human (or humanized) antigen binding protein, for example, a monoclonal antibody, which induces FGF21 type signaling, for example, an agonist antibody that mimics FGF21 function. Such an antibody is a molecule with FGF21-like activity and selectivity, but with therapeutically desirable added characteristics typical for an antibody such as, for example, protein stability, absence of immunogenicity, ease of production and long in vivo half-life.

SUMMARY OF THE INVENTION

An isolated antigen-binding protein that induces FGF21-mediated signaling is provided. An isolated antigen-binding protein is also provided that specifically binds to at least one of: (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; and (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3C and FGFR4, wherein the antigen binding protein induces FGF21-mediated signaling.

In one embodiment, the provided antigen binding proteins comprise an amino acid sequence selected from the group consisting of: (a) a light chain CDR3 comprising a sequence selected from the group consisting of: (i) a CDR3 sequence of light chain that differs by at most a total of three amino acid additions, substitutions and / or deletions from a CDR3 sequence selected from the group consisting of the L1-L18, IDS light chain CDR3 sequences. SEQ. ^Nos: 180-194; (ii) QVWDXxXaSDHW (SEQ ID. NO: 276); (Üi) QQX ₃ GX ₄ X ₅ X6X7T (SEQ ID. NO: 283); (iv) LQHNSYPLT (ID. DE

SEQ. No.: 267); (v) MQSLQTPFT (SEQ ID NO: 268); (saw)

QQYNNWPPT (SEQ ID. NO: 269); (vii) MQSIQLPRT (SEQ ID NO: 270); (viíí) QQANDFPIT (SEQ ID NO: 271); (ix) MQALQTPCS (SEQ ID. NO: 272); (b) a heavy chain CDR3 sequence comprising a sequence selected from the group consisting of: (i) a heavy chain CDR3 sequence that differs by a maximum of four additions, substitutions and / or deletions from amino acids from a CDR3 sequence selected from the group consisting of the H1-H18 heavy chain CDR3 sequences, IDS. SEQ. ^Nos: 145-157; (Ü) X ₃₄ Xi ₆ Xi7Xi8GXi ₉ YYYX2oGMDV (SEQ ID. NO:

322); (Üi) SLIWX ₂ iVYX22LDX ₂ 3 (SEQ ID. NO: 326); (iv) IVWPAAIQSYYYYYGMGV (SEQ ID. NO: 311); (v)

DPDGDYYYYGMDV (SEQ ID. NO: 312); (vi) TYSSGWYVWDYYGMDV (SEQ ID. NO: 313); (vii) DRVLSYYAMAV (SEQ ID NO: 314); (viii) VRIAGDYYYYYGMDV (SEQ ID. NO: 315); (ix) ENIWIPAAIFAGWFDP (SEQ ID. NO: 316); and (x)

DRAAAGLHYYYGMDV (SEQ ID. NO: 317); or (c) the light chain CDR3 sequence from (a) and the heavy chain CDR3 sequence from (b); where X _x is G, S or N; X ₂ is N, I am

T; X ₃ is C, Y or S; X ₄ is G OR S; X ₅ is A OR S; X ₆ is P or F; X is L or absent; X ₃₄ is I, V or S; X ₁₆ is L or V; X ₁₇ is L, Tou

V; X ₁₈ is L, V, G or T; X _i9 is A, G or absent; X ₂ o is Y, C or D; X ₂ 1 is I or M; X ₂₂ is A or V; and X ₂ 3 is H or Y; and where the antigen binding protein specifically binds to:

(i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2C, FGFR3C and FGFR4.

In another embodiment, the supplied antigen binding proteins comprise: (a) a sequence of light chain CDR1 selected from the group consisting of: (i) a light chain CDR1 that differs by a maximum of three additions, substitutions and / or deletions of amino acids from a CDR1 sequence of L1-L18, IDS. SEQ. ^Nos: 158-17 0;

(ii) RASQX9X10X11X12X13X14LA (SEQ ID. NO: 304); (iii)

GGNNIGSX ₁₅ SVH (SEQ ID NO: 307); (iv)

RSSQSLLX29X30NGX31X32X33LD (SEQ ID. N °: 310); (v)

RASQSVNSNLA (SEQ ID. NO: 295); (vi) RASQDIRYDLG (SEQ ID. NO: 296); (vii) RASQGISIWLA (SEQ ID NO: 297); and (viii) KSSQSLLQSDGKTYLY (SEQ ID. NO: 298); (b) a light chain CDR2 sequence selected from the group consisting of: (i) a light chain CDR2 that differs by at most two additions, substitutions and / or deletions from

amino acids from an sequence from CDR2 from I ..1-L18, IDS. IN SEQ. N ^os : 171-179; (ii) LGSX ₂ ^ iRAS (ID . IN SEQ. No. : 290); (ÍÜ) GX; ₈ SX ₂₈ RAT (ID. SEQ. Ν ' ⁰ : 2 94); (iv) AASSLQS (ID. OF SEQ. N °: 2 84); (v) GVSTRAT ( ID. IN SEQ. N °: 285 ); (saw) DDSDRPS (ID. OF SEQ . N °: 286) ; (vii) EVSNRFS (ID. SEQ. N °: 287) ); (O an sequence of CDR1 of jail heavy

selected from the group consisting of: (i) a heavy chain CDR1 that differs by at most two amino acid additions, substitutions and / or deletions from a CDR1 sequence of H1-H18, IDS. SEQ. ^Nos: 121-131; and (ii)

NARMGVX39 (SEQ ID. NO: 352); (iii) X ₄₀ YGIH (SEQ ID NO: 355); (iv) DLSMH (SEQ ID NO: 345); (v) DAWMS (SEQ ID. NO .: 346); (vi) TYAMS (SEQ ID. NO .: 347); (vii) SYFWS (SEQ ID. NO .: 348); (viii) SYYWS (SEQ ID. NO .: 131); (ix) SGGYNWS (SEQ ID. NO: 349); (d) a heavy chain CDR2 selected from the group consisting of: (i) a heavy sequence that differs by at most three amino acid additions, substitutions and / or deletions from a sequence

of H1-H18 CDR2 , IDS IN SEQ. ^Nos: 132-144; (ii) HI FSNDEKS YSTSLKX ₂₄ (ID. IN SEQ. N °: 333); (iii) x ₂₅ isgsgvstx ₂₆ yadsvkg (ID. IN SEQ. N °: 338); (iv) viwydgsx ₃₅ kyyx _3S dsvkg (ID. IN SEQ. No. : 341); (v) X ₃₇ IYX ₃₈ SGSTX ₄ iYNPSLKS (ID. IN SEQ. N °: 344); (Saw) GFDPEDGETIYAQKFQG (ID. IN SEQ. N °: 327); (vii) RIKSKTDGGTTDYAAPVKG (ID. IN SEQ. N °: 328); (viii) RIYTSGSTNYNPSLKS ( ID. IN SEQ. N °: 329); (ix) RIKSKDGGTTDYAAPVKG (ID. IN SEQ. N °: 330); (x)

RIKSKX ₄₂ DGGTTDYAAPVKG (SEQ ID. NO: 483); where X ₉ is N OR S; X ₁₀ is V or F; Xn is D OR S; X ₃₂ is G or S; X ₁₃ is S, Nou

T; X ₁₄ is S OR Y; X ₁₅ is E or Q; X ₂₉ is Y OR H; X ₃₀ is Y OR S; X is F or Y; X ₃₂ is T or N; X ₃₃ is Y or F; X ₂₇ is N or D; X ₈ is A or T; X ₂₈ is S or F; X ₃₉ is Y or N; X ₂₄ is Y or N; X ₂₅ is G OR A; X is Η, Y or N; X ₃₅ is D or I; X ₃₆ is A or G; X ₃₇ is N or R; X ₃₈ is

Y or T; X ₄ i is Y or N; X ₄₂ is T or absent; (e) the light chain CDR1 of (a) and the light chain CDR2 of (b); (f) the light chain CDR1 of (a) and the heavy chain CDR1 of (c); (g) the light chain CDR1 of (a) and the heavy chain CDR2 of (d); (h) the light chain CDR1 (b) and the heavy chain CDR1 of (c); (i) the heavy chain CDR1 of (c) and the heavy chain CDR2 of (d); (j) the light chain CDR2 of (b) and the heavy chain CDR2 of (d); (k) the light chain CDR1 of (a), the light chain CDR2 of (b) and the heavy chain CDR1 of (c); (1) the light chain CDR2 of (b), the heavy chain CDR1 of (c) and the heavy chain CDR2 of (d); (m) the light chain CDR1 of (a), the heavy chain CDR1 of (c) and the heavy chain CDR2 of (d); or (n) the light chain CDR1 of (a), the light chain CDR2 of (b), the

Heavy chain CDR2 of (c) and heavy chain CDR2 of (d), wherein said antigen binding protein specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4.

In yet another embodiment, the provided antigen binding proteins comprise: (a) a light chain variable domain comprising: (i) a light chain CDR1 sequence selected from the IDS. SEQ. ^Nos: 158-170; (ii) a light chain CDR2 sequence selected from the IDS. SEQ. ^Nos: 171-179; (iii) a light chain CDR3 sequence selected from the IDS. SEQ. No. ^s : 180-194; and (b) a heavy chain variable domain comprising: (i) a heavy chain CDR1 sequence selected from the IDS. SEQ. ^Nos: 121-131; (ii) a heavy chain CDR2 sequence selected from the IDS. SEQ. ^Nos: 132-144; and (iii) a heavy chain CDR3 sequence selected from the IDS. SEQ. ^Nos: 145-157; or (c) the light chain variable domain of (a) and the heavy chain variable domain of (b), wherein the antigen binding protein specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3C and FGFR4.

In an additional embodiment, the provided antigen binding proteins comprise: (a) a sequence of the variable domain of the light chain selected from the group consisting of: (i) amino acids that have a sequence at least 80% identical to a sequence of the domain light chain variable selected from V _L 1-V _L 18, IDS. SEQ. ^Nos: 4865; (ii) an amino acid sequence encoded by a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence that encodes the V _L 1-V _L 18, IDS light chain variable domain sequence. SEQ. No. ^s : 48-65; (b) a sequence of the variable domain of the heavy chain selected from the group consisting of: (i) a sequence of amino acids that is at least 80% identical to a sequence of the variable domain of the heavy chain of V _H 1 -V _H 18 of IDS. SEQ. ^Nos: 66-84; (ii) an amino acid sequence encoded by a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence that encodes the variable domain sequence of the V _H 1-V _H 18, IDS heavy chain. SEQ. ^Nos: 66-84; or (c) the light chain variable domain of (a) and the heavy chain variable domain of (b); wherein the antigen binding protein specifically binds to: (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. In particular embodiments, the provided antigen binding proteins comprise: (a) a sequence of the variable domain of the light chain selected from the group consisting of: V _L 1-V _L 18 of the IDS. SEQ. ^Nos: 48-65; (b) a sequence of the variable domain of the heavy chain selected from the group consisting of: V _H 1V _h 18 of the IDS. SEQ. No. ^s : 66-84; or (c) the light chain variable domain of (a) and the heavy chain variable domain of (b), wherein the antigen binding protein specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. In other particular embodiments, the supplied antigen binding proteins, the light chain variable domain and a heavy chain variable domain are selected from the group of combinations consisting of: V _L 1V _H 1, V _L 2V _H 2, V _L 3V _H 3, V _L 3V _H 4, V _l 4V _h 5, V _l 5V _h 6, V _l 6V _h 7, V _L 7V _H 8, V _L 8V _H 8, V _L 9V _H 9, V _L 9V _H 10, V _L 10V _H ll, V _l 11V _h 11, V _l 12V _h 12, V _L 13V _H 13, V _L 14V _H 14, V _L 15V _H 15,

V _L 16V _H 16, V _l 17V _h 17 and V _L 18V _H 18, where the antigen binding protein specifically binds to: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. In yet other additional embodiments, the provided antigen binding proteins further comprise: (a) the constant sequence of the ID light chain. SEQ. No. 10; (b) the constant sequence of the ID light chain. SEQ. No.: 11; (c) the constant sequence of the ID heavy chain. IN

SEQ. No.: 9; or (d) the constant sequence of the ID light chain. SEQ. No. 10 or ID. SEQ. No. 11 and the constant sequence of the ID heavy chain. SEQ. N °: 9.

The provided antigen binding proteins can take many forms and can be, for example, a human antibody, a humanized antibody, a chimeric antibody, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, an antigen binding antibody fragment , a single chain antibody, a diabody, a triabody, a tetrabody, a Fab fragment, an F (fab ') 2 fragment, a domain antibody, an IgD antibody, an IgE antibody, an IgM antibody, an IgGl antibody, an IgG2 antibody, an IgG3 antibody, an IgG4 antibody or an IgG4 antibody that have at least one mutation in the hinge region.

In another embodiment, the antigen binding proteins provided when linked to: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4: (a) bind to: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, with substantially the same Kd as a reference antibody; (b) induce FGF21 type signaling of 10% or more than signaling induced by a wild type FGF21 pattern comprising the mature form of the ID. SEQ. No. 2, as measured in an ELK-luciferase reporter assay; (c) exhibit an EC50 of 10 nM or less of FGF21 type signaling in an assay selected from the group consisting of: (i) an in vitro assay based on recombinant cell and mediated by FGFRlc / β-Klotho; and (ii) a functional in vitro human adipocyte assay; (d) exhibit an EC50 of less than 10 nM agonist activity in FGFRlc in the presence of βKlotho in an in vitro reporter assay mediated by recombinant FGFRlc receptor; and (e) exhibit an EC50 greater than 1 pM of FGFRlc agonist activity in the absence of βKlotho in an in vitro reporter assay mediated by recombinant FGFRlc receptor; or (f) compete for binding with a reference antibody for: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, wherein the reference antibody comprises a combination of sequences from the variable domain of the light chain and the heavy chain selected from the group consisting of V _L 1V _H 1, V _l 2V _h 2, V _l 3V _h 3, V _l 3V _h 4, V _l 4V _h 5, V _L 5V _H 6, V _L 6V _H 7, V _L 7V _H 8, V _L 8V _H 8,

V _l 9V _h 9, V _l 9V _h 10, V _l 10V _h 11, V _l 11V _h 11, V _l 12V _h 12, V _l 13V _h 13,

V _l 14V _h 14, V _l 15V _h 15, V _l 16V _h 16, V _l 17V _h 17 and V _L 18V _H 18. In other embodiments, the supplied antigen binding proteins can, when bound to (i) β -Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4: (a) reducing blood glucose in an animal model; (b) to reduce serum lipid levels in an animal model; (c) reducing insulin levels in an animal model; or (d) two or more of (a) and (b) and (c).

In specific embodiments, the provided antigen binding proteins comprise: (a) a heavy chain comprising one of the IDS. SEQ. ^Nos: 31, 32, 390-401, 4 04405; (b) a light chain comprising one of the IDS. SEQ. ^Nos: 13, 14, 385-389, 402-403; or (c) a combination comprising a heavy chain of (a) and a light chain of (b).

Antigen binding proteins are also provided that are capable of binding to human wild-type β-Klotho (SEQ ID. NO: 7), but which do not bind to a chimeric form of β-Klotho, in which the chimeric form of βKlotho comprises a human β-Klotho framework, in which murine β-Klotho sequences replace wild-type human residues in at least one of (a) positions 1-80; (b) positions 303-522; (c) positions 852-1,044; and (d) combinations of these.

In another aspect, the present disclosure provides antigen-binding proteins that are capable of binding to human wild-type β-Klotho (SEQ ID NO:

7) in at least one of (a) positions 1-80; (b) positions 303-522; (c) positions 852-1,044; and (d) combinations of these.

In yet another aspect, the present disclosure provides antigen binding proteins that are capable of competing with an antigen binding protein of claims 8 or 13 for binding to human wild-type βKlotho residues in at least one of (a) positions 1-80; (b) positions 303-522; (c) positions 8521,044; and (d) combinations of these.

Also provided is a pharmaceutical composition comprising one or more antigen binding proteins provided herein, mixed with a pharmaceutically acceptable carrier thereof.

In a further aspect, isolated nucleic acid molecules are also provided that encode the antigen binding proteins disclosed herein. In some cases, isolated nucleic acid molecules are operationally linked to a control sequence. In embodiments, these nucleic acids comprise a polynucleotide sequence that encodes the variable domain of the light chain, the variable domain of the heavy chain, or both, of an antigen binding protein provided herein. In particular embodiments, the nucleic acids comprise: (a) _{VL 1-VL} 18 (SEQ IDs ^Nos:.. 65 4825); (b) V _H 1-V _H 18 (SEQ IDs ^Nos:.. 66-84); or (c) one or more strings of (a) and one or more strings of (b).

In another aspect, expression vectors and host cells transformed or transfected with the expression vectors comprising the previously mentioned isolated nucleic acid molecules encoding the antigen binding proteins disclosed herein are also provided.

In another aspect, methods are also provided for preparing antigen binding proteins that specifically or selectively bind to: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 and comprising the step of preparing the antigen-binding protein by a host cell that secretes the antigen-binding protein.

Other modalities provide a method of preventing or treating a condition in an individual in need of such treatment, which comprises administering a therapeutically effective amount of a pharmaceutical composition provided herein to an individual, wherein the condition is treatable by reducing blood glucose, insulin or serum lipid levels. In modalities, the condition is type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease or metabolic syndrome.

These and other aspects are described in more detail here. Each of the aspects provided can encompass several modalities provided here. Therefore, it is envisaged that each of the modalities involving an element or combinations of elements can be included in each aspect described, and all these combinations of the aspects and modalities above are expressly considered. Other characteristics, objects and advantages of the disclosed antigen binding proteins and associated methods and compositions are apparent in the detailed description that will be presented below.

BRIEF DESCRIPTION OF THE DRAWINGS

Figures 1A-IB are an alignment showing the sequence homology between human FGFRlc (GenBank Access Ν 'P11362; SEQ ID NO: 356) and murine FGFRlc (GenBank Access NP_034336; ID. SEQ NO: 357);

several features are highlighted, including the signal peptide, transmembrane sequence, heparin binding region, and a consensus sequence (SEQ ID. NO: 358) is provided.

Figures 2A-2C are an alignment showing the sequence homology between human β-Klotho (GenBank Accession No. NP_783864; SEQ ID. NO .: 359) and murine β-Klotho (GenBank Accession Ν ' NP_112457; SEQ ID NO:

360); several characteristics are highlighted, including the transmembrane sequence and two domains of glycosyl hydrolase, and a consensus sequence (SEQ ID. N °:

361) is provided.

Figure 3 is a flow cytometry profile of cells stained with FGF21-Alexa 647 that were used as an immunogen to generate antigen binding proteins; the figure shows the level of expression of a FGF21R (a complex comprising FGFRlc and β-Klotho) and the binding to FGF21.

Figure 4 is a sequence (SEQ ID. NO: 362) showing an Fc fusion protein that was used as an immunogen to generate antigen-binding proteins; the immunogen comprises the extracellular domain (ECD) of human FGFRlc fused to an IgGl Fc by means of a Gly5 linker (SEQ ID. NO: 379); the FGFRlc component is in uppercase, the linker is in italics and underlined, and the Fc is in lowercase.

Figure 5 is a sequence (SEQ ID. NO: 363) showing an Fc fusion protein that was used as an immunogen to generate antigen-binding proteins; the immunogen comprises the extracellular domain (ECD) of human βKlotho fused to an IgGl Fc by means of a Gly5 linker (SEQ ID. NO: 379); the β-Klotho component is in uppercase letters, the linker is in italics and underlined, and the Fc is in lowercase letters.

Figure 6 is an SDS-PAGE gel showing the level of purity obtained by preparations of a soluble FGF21 receptor complex comprising FGFRlc ECD-Fc and β-Klotho ECD-Fc, which was employed as an immunogen for generate antigen binding proteins.

Figure 7 is a series of graphs generated by an ELK-luciferase reporter assay as described herein performed on the recombinant CHO clone 2E10, demonstrating the ability of some of the antigen binding proteins to induce FGF21-like signaling in recombinant CHO cells. that express a FGF21 receptor complex comprising FGFRlc and β-Klotho.

Figure 8 is a series of graphs generated by an ERK1 / 2 phosphorylation assay as described herein, demonstrating the ability of some of the antigen binding proteins to induce FGF21-like signaling in rat L6 cells. The X axis is the concentrations of the antigen binding proteins and the Y axis is the percentage of total ERK1 / 2 phosphorylated ERK1 / 2.

Figure 9 is a series of graphs generated by an ERK1 / 2 phosphorylation assay as described herein, demonstrating that the FGF21 type signaling mediated by the antigen binding protein in L6 cells is FGFRlc / βKlotho-specific.

Figure 10 is a series of graphs generated by an ERK phosphorylation assay as described herein, demonstrating that some antigen binding proteins are capable of inducing FGF21-like signaling in human adipocyte cells.

Figure 11a is a series of binding sensograms (response units versus time) demonstrating that some of the antigen binding proteins that induce FGF21-mediated signaling bind to human β-Klotho at two different but partially overlapping binding sites, represented by 24H11 (Group A) and 17D8 (Group B), while antigen-binding proteins that do not induce FGF21-mediated signaling (2G10, 1A2) do not bind to these sites.

Figure 11b is a series of binding sensograms (response units versus time) that demonstrate a third binding site in human β-Klotho that has been identified for Group C antigen binding proteins represented by 39F7.

Figure 12 is a series of binding sensograms (response units versus time) demonstrating that some of the antigen binding proteins (12E4, 24H11, 17C3, 18B11) that induce FGF21-mediated signaling interfere with β-Klotho binding to FGF21 , while other antigen-binding proteins (21H2, 17D8, 18G1) do not.

Figure 13 is an alignment of the variable regions of some of the antigen binding proteins that have been generated; the framework and CDR regions are identified.

Figure 13 reveals IDS. SEQ. ^Nos: 364, 59, 365, 60, 366,

61, 367, 62, 368, 57, 369, , 55 , 51-52, 56, 56, 53-54 63 -65 370, , 58, 371, , 50, 50, 49, 48, 372 , 78, 373 66 -69, 79, 374 76, 81, 375, 70, 73, 73, 71- 72, 376, 83, 82, 84, 377, 80

378, 75 and 74, respectively, in order of occurrence.

Figure 14 is a diagram that graphically illustrates the study design for a 68-day study performed on obese Cynomolgus monkeys.

Figure 15 is a graph describing the effects of vehicle and 16H7 on food intake in the morning meal of the obese Cynomolgus monkeys studied.

Figure 16 consists of two graphs that describe the effects of vehicle and 16H7 on fruit and food intake in the afternoon of the obese Cynomolgus monkeys studied 20.

Figure 17 is a graph describing the effects of vehicle and 16H7 on the body weight of the obese Cynomolgus monkeys studied.

Figure 18 is a graph showing the effects of vehicle and 16H7 on the body mass index (BMI) of the obese Cynomolgus monkeys studied.

Figure 19 is a graph showing the effects of vehicle on the abdominal circumference (AC) of the obese Cynomolgus monkeys studied.

Figure 20 is a graph showing the effects of vehicle and 16H7 on the skin fold thickness (FTS) of the obese Cynomolgus monkeys studied.

Figure 21 is a graph showing the effects of vehicle and 16H7 on glucose levels during glucose tolerance tests of the obese Cynomolgus monkeys studied.

Figure 22 is a graph showing the effects of vehicle and 16H7 on plasma insulin levels during glucose tolerance tests of the obese Cynomolgus monkeys studied.

Figure 23 is a graph showing the effects of vehicle and 16H7 on fasting blood glucose levels of the obese Cynomolgus monkeys studied.

Figure 24 is a graph showing the effects of vehicle and 16H7 on plasma insulin fasting levels of the obese Cynomolgus monkeys studied.

Figure 25 is a graph showing the effects of vehicle and 16H7 on the blood glucose levels of the obese Cynomolgus monkeys studied.

Figure 26 is a graph showing the effects of vehicle and 16H7 on plasma insulin levels of the obese Cynomolgus monkeys studied.

Figure 27 is a graph showing the effects of vehicle and 16H7 on plasma fasting levels of triglycerides of the obese Cynomolgus monkeys studied.

Figure 28 is a graph showing the effects of vehicle and 16H7 on the plasma levels of triglycerides in the obese Cynomolgus monkeys studied.

Figure 29 is a representation schematic what describes chimeras in β-Klotho human-de mouse what Were build and used for study of the call in

antigen binding proteins.

Ά Figure 30 is a schematic representation that describes the mouse human-β-Klotho chimeras that were constructed, and also includes qualitative binding data for FGF21, 16H7, 37D3 and 39F7.

Figures 31A-C consist of a series of graphs that describe connection data for eight of the 16H7 and 22H5 variants that were constructed, as well as for 22H5 and 16H7.

Figures 32A-C consist of a series of graphs that describe the results of ELISA assays that were used to demonstrate that several of the 22H5 and 16H7 variants have binding ability.

Figure 33 is a bar graph that compares offrates for several variants of 22H5 and 17H7 that were generated.

Figure 34 consists of two graphs that describe the binding curves for 39F11 when titrated with FGF21 and for FGF21 when titrated with 39F11; the graphics demonstrate an additive effect.

Figure 35 consists of two graphs that describe the binding curves for 16H7 when titrated with 39F11 and 39F11 when titrated with 16H7; the graphics demonstrate an additive effect.

DETAILED DESCRIPTION OF THE INVENTION

The section headings used here are for organizational purposes only and should not be considered as limiting the theme described.

Unless otherwise defined herein, the scientific and technical terms used in connection with this application have the meanings that are commonly implied by those skilled in the art. In addition, unless needed differently by context, terms in the singular must include pluralities and terms in the plural must include the singular.

Generally, the nomenclatures used in relation to the techniques of cell and tissue culture, molecular biology, immunology, microbiology, genetics and chemistry and hybridization of proteins and nucleic acids described herein are well known and commonly used in the art. The methods and techniques of the present application are generally carried out according to conventional methods well known in the art and as described in several general and more specific references which are cited and discussed throughout the present specification, unless otherwise indicated. See, e.g., Sambrook et al, Molecular Cloning:.. A Laboratory Manual, 3rd Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (2001), Ausubel et al, Current Protocols in Molecular Biology, Greene Publishing Associates (1992), and Harlow and Lane Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1990), which are hereby incorporated by reference. Enzymatic reactions and purification techniques are carried out according to the manufacturer's specifications, as commonly obtained in the technique or as described here. The terminology used and laboratory procedures and techniques of analytical chemistry, synthetic organic chemistry and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques can be used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and release, and patient treatment.

It should be understood that the present disclosure is not limited to the particular methodology, protocols and reagents etc. described herein to the extent that they may vary. The terminology used here is intended only to describe particular modalities, and is not intended to limit the scope of the present disclosure.

Except in the operational examples, or when indicated differently, all numbers that express quantities of ingredients or reaction conditions used herein must be understood as modified in all cases by the term about. The term about, when used in connection with percentages, can mean ± 5%, for example, 1%, 2%, 3% or 4%.

I. DEFINITIONS

As used herein, the terms one and one mean one or more, unless specifically stated differently.

An antigen binding protein is a protein that comprises a portion that binds to an antigen or target and, optionally, a frame or framework portion that allows the antigen binding portion to adopt a conformation that promotes the binding of the antigen protein. antigen binding to antigen. Examples of antigen binding proteins include a human antibody, a humanized antibody; a chimeric antibody; a recombinant antibody; a single chain antibody; a diabody; a triabody; a tetrabody; a Fab fragment; an F (ab ') ₂ fragment; an IgD antibody; an IgE antibody; an IgM antibody; an IgGl antibody; an IgG2 antibody; an IgG3 antibody; or an IgG4 antibody, and fragments thereof. The antigen-binding protein may comprise, for example, an alternative protein framework or an artificial framework with grafted CDRs or CDR derivatives. These frameworks include, without limitation, antibody derived frameworks that comprise mutations introduced, for example, to stabilize the three-dimensional structure of the antigen binding protein, in addition to fully synthetic frameworks that comprise, for example, a biocompatible polymer. See, for example, Korndorfer et al, 2003, Proteins: Structure, Function, and Bioinformatics, 53 (1): 121-129 (2003); Roque et al, Biotechnol. Prog. 20: 639-654 (2004). In addition, peptide antibody mimetics (PAMs) can be used, in addition to frameworks based on antibody mimetics that use fibronectin components as a framework.

An antigen-binding protein can have, for example, the structure of a naturally occurring immunoglobulin. An immunoglobulin is a tetrameric molecule. In a naturally occurring immunoglobulin, each tetramer is composed of two identical pairs of polypeptide chains, each pair having a light chain (about 25 kDa) and a heavy chain (about 50-70 kDa). The amino terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The carboxy terminal portion of each chain defines a constant region primarily responsible for the effector function. Human light chains are classified as kappa and lambda light chains. Heavy chains are classified as mu, delta, gamma, alpha or epsilon, and define the antibody isotype as IgM, IgD, IgG, IgA and IgE, respectively. Within the light and heavy chains, the variable and constant regions are joined by a J region of about 12 or more amino acids, with the heavy chain also including a D region of about 10 more amino acids. See generally, Fundamental Immunology, Chapter 7 (Paul, W., ed., 2nd Edition , Raven Press, NY

(1989)) (incorporated by reference in its entirety for all purposes). The variable regions of each light / heavy chain pair form the antibody binding site, such that an intact immunoglobulin has two binding sites.

Naturally occurring immunoglobulin chains exhibit the same general structure as relatively conserved framework regions (FR) joined by three hypervariable regions, also called complementarity determining regions or CDRs. From terminal N to terminal C, both light and heavy chains comprise the FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4 domains. The assignment of amino acids to each domain can be made according to Kabat et al settings in Sequences of Proteins of Immunological Interest, 5th Edition, US Dept, of Health and

Human Services, PHS, NIH, NIH Publication No. 91-3242, 1991. As desired, CDRs can also be redefined according to an alternative nomenclature scheme, for example, that of Chothia (see Chothia and Lesk, 1987, J Mol. Biol. 196: 901-917; Chothia et al., 1989, Nature 342: 878-883 or Honegger and Pluckthun, 2001, J. Mol. Biol. 309: 657-670).

In the context of the present disclosure, an antigen binding protein is said to specifically bind or selectively bind to its target antigen when the dissociation constant (K _D ) is <10 ^-8 Μ. The antibody specifically binds to the antigen with high affinity when the KD is <5 x 10 ' ⁹ M, and with very high affinity when the KD is <5 x 10 ^{_ 1} ° M. In one embodiment, the antibodies will bind to FGFRlc, β-Klotho, either to FGFRlc or β-Klotho, or to a complex comprising FGFRlc and β-Klotho, including human FGFRlc, human β-Klotho, or both human FGFRlc and human β-Klotho, with a K _D between about IO ' ⁷ M and IO' ¹² M and, in yet another modality, the antibodies will bind with a K ^D <5 x IO ' ⁹ .

An antibody refers to an intact immunoglobulin or antigen binding portion of it that competes with the intact antibody for specific binding, unless otherwise specified. Antigen binding moieties can be produced by recombinant DNA techniques or by enzymatic or chemical dividing of intact antibodies. Antigen binding moieties include, inter alia, Fab, Fab ', F (ab') ₂ , Fv, domain antibodies (dAbs), fragments that include complementarity determining regions (CDRs), single chain antibodies (scFv), chimeric antibodies, diabodies, triabodies, tetrabodies and polypeptides that contain at least a portion of an immunoglobulin that is sufficient to confer specific antigen binding to the polypeptide.

A Fab fragment is a monovalent fragment that has the domains of V _L , V _H , C _L and C _H 1; an F (ab ') ₂ fragment is a divalent fragment that has two Fab fragments linked by a disulfide bridge in the hinge region; an Fd fragment has the V _H and C _H 1 domains; an Fv fragment has the V _L and V _H domains of a single arm of an antibody; and a dAb fragment has a domain V _H, V _L domain, or an antigen-binding fragment of a V _H domain or V _L (US Patents ^Nos 6,846,634, 6,696,245, US Application Publication ^Nos 05/0202512, 04/0202995, 04/0038291, 04/0009507, 03/0039958, Ward et al, Nature 341:. 544-546 (1989)).

A single chain antibody (scFv) is an antibody in which a region of V _L and one region of V _H are joined by means of a linker (for example, a synthetic sequence of amino acid residues) to form a continuous protein chain in that the linker is long enough to allow the protein chain to fold over itself and form a monovalent antigen binding site (see, for example, Bird et al., Science 242: 423-26 (1988) and Huston and cols., 1988, Proc. Natl. Acad. Sci. USA 85: 5.87983 (1988)). Diabodies are bivalent antibodies that comprise two polypeptide chains, where each polypeptide chain comprises domains of V _H and V _L joined by a linker that is too short to allow pairing between two domains on the same chain, thus allowing each domain pair with a complementary domain in another polypeptide chain (see, for example, Holliger et al., 1993, Proc. Natl. Acad. Sci. USA 90: 6.444-48 (1993), and Poljak et al., Structure 2: 1.121 -23 (1994)). If the two polypeptide chains of a diabody are identical, then a diabody resulting from their pairing will have two identical antigen binding sites. Polypeptide chains that have different sequences can be used to produce a diabody with two different antigen binding sites. Similarly, tribodies and tetrabodies are antibodies that comprise three and four polypeptide chains, respectively, and that form three and four antigen binding sites, respectively, that can be the same or different.

Complementarity determining regions (CDRs) and framework regions (FR) of a certain antibody can be identified using the system described by Kabat et al. in Sequences of Proteins of Immunological Interest, 5th edition, US Dept, of Health and Human Services, PHS, NIH, NIH Publication No. 91-3242, 1991. As desired, the CDRs can also be redefined according to a schema alternative nomenclature, for example, that of Chothia (see Chothia and Lesk, 1987, J. Mol. Biol. 196: 901-917; Chothia et al., 1989, Nature 342: 878-883 or Honegger and Pluckthun, 2001, J Mol. Biol. 309: 657-670). One or more CDRs can be incorporated into a molecule either covalently or non-covalently to make it an antigen-binding protein. An antigen-binding protein can incorporate the CDR (s) as part of a larger polypeptide chain, can covalently attach the CDR (s) to another polypeptide chain, or can incorporate the CDR (s) ) not covalently. CDRs allow the antigen-binding protein to specifically bind to an antigen of interest in particular.

An antigen binding protein can have one or more binding sites. If there is more than one connection site, the connection sites can be identical to each other or they can be different. For example, a naturally occurring human immunoglobulin typically has two identical binding sites, while a bispecific or bifunctional antibody has two different binding sites.

The term human antibody includes all antibodies that have one or more variable and constant regions derived from human immunoglobulin sequences. In one embodiment, all variable and constant domains are derived from human immunoglobulin sequences (a fully human antibody). These antibodies can be prepared in a variety of ways, examples of which are described below, including by immunizing with an antigen of interest to a mouse that is genetically modified to express antibodies derived from genes encoding human heavy and / or light chain, for example , a mouse derived from a Xenomouse®, UltiMab ™ or Velocimmune® system. Phage-based approaches can also be employed.

A humanized antibody has a sequence that differs from the sequence of an antibody derived from a non-human species by one or more amino acid substitutions, deletions and / or additions, such that the humanized antibody is less likely to induce an immune response and / or induce a less severe immune response when compared to the antibody of a non-human species when administered to a human individual. In one embodiment, certain amino acids in the framework and constant domains of the heavy and / or light chains of the antibody of the non-human species are mutated to produce the humanized antibody. In another embodiment, the constant domain (s) of a human antibody are fused to the variable domain (s) of a non-human species. In another embodiment, one or more amino acid residues in one or more CDR sequences of a non-human antibody are altered to reduce the likely immunogenicity of the non-human antibody when it is administered to a human individual, in which the altered amino acid residues are not are crucial for immunospecific binding of the antibody to its antigen, or the changes in the amino acid sequence that are made are conservative changes, such that the binding of the humanized antibody to the antigen is not significantly worse than the binding of the non-human antibody to the antigen . Examples of how to make humanized antibodies may be found in US Patents ^Nos 6,054,297, 5,886,152 and 5,877,293.

The term chimeric antibody refers to an antibody that contains one or more regions of an antibody and one or more regions of one or more other antibodies. In one embodiment, one or more of the CDRs are derived from a human antibody that binds to: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. In another embodiment, all CDRs are derived from a human antibody that binds to: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. In another embodiment, the CDRs of more than one human antibody that bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 are mixed and combined into a chimeric antibody. For example, a chimeric antibody may comprise a light chain CDR1 of a first human antibody that binds to: (i) βKlotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, CDR2, and a light chain CDR3 of a second human antibody that binds to: (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, and the heavy chain CDRs of a third antibody that binds to: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. In addition, framework regions can be derived from one of the same antibodies that bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, of one or more different antibodies, for example, a human antibody, or a humanized antibody. In an example of a chimeric antibody, a portion of the heavy and / or light chain is identical, homologous or derived from an antibody of a particular species or that belongs to a particular antibody class or subclass, while the rest of the (s) chain (s) is identical, homologous or derived from an antibody or antibodies of another species or that belongs to another class or subclass of antibody. Also included are fragments of these antibodies that exhibit the desired biological activity (for example, the ability to specifically bind to: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex that comprises β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4).

The term light chain includes a full-length light chain and fragments of it that have enough variable region sequence to confer binding specificity. A full-length light chain includes a variable region domain, V _L , and a constant region domain, C _L. The light chain variable region domain is at the amino terminus of the polypeptide. Light chains include kappa (K) and lambda (λ) chains.

The term heavy chain includes a full-length heavy chain and fragments thereof that have sufficient variable region sequence to confer specificity

binding. A full-length heavy chain includes a domain of variable region, V _H , it's three domains of constant region , C _H 1, C _h 2 and C _h 3. 0 domain of V _H is at the terminal amino do polypeptide, and the domains of CH They are at the terminal

carboxyl, with C _H 3 being closer to the carboxy terminal of the polypeptide. Heavy chains can be of any isotype, including IgG (including IgGl, IgG2, IgG3 and IgG4 subtypes), IgA (including IgAl and IgA2 subtypes), IgM and IgE.

The term immunologically functional fragment (or simply fragment) of an antigen binding protein, for example, an antibody or immunoglobulin chain (heavy or light chain), as used herein, is an antigen binding protein that comprises a portion ( regardless of how that portion is obtained or synthesized) of an antibody lacking at least some of the amino acids present in a full-length chain, but which is capable of specifically binding to an antigen. These fragments are biologically active in that they specifically bind to the target antigen and can compete with other antigen binding proteins, including intact antibodies, for specific binding to a certain epitope. In one aspect, such a fragment will retain at least one CDR present in the full-length light or heavy chain and, in some embodiments, will comprise a single heavy chain and / or light chain or portion thereof. These biologically active fragments can be produced by recombinant DNA techniques, or they can be produced by enzymatic or chemical dividing of antigen binding proteins, including intact antibodies. Immunologically functional immunoglobulin fragments include, without limitation, Fab, Fab ', F (ab') ₂ , Fv, domain antibodies and single chain antibodies, and can be derived from any mammalian source, including, without limitation, humans, mouse, rat, camelid or rabbit. It is further contemplated that a functional portion of the antigen-binding proteins disclosed herein, for example, one or more CDRs, could be covalently linked to a second protein or small molecule to create a therapeutic agent directed at a particular target in the body, that has bifunctional therapeutic properties, or that has a prolonged serum half-life.

An Fc region contains two heavy chain fragments that comprise the C _H 2 and C _H 3 domains of an antibody. The two heavy chain fragments are held together by two or more disulfide bonds and by hydrophobic interactions of the C _H 3 domains.

A Fab 'fragment contains a light chain and a portion of a heavy chain that contains the V _H domain and the C _H 1 domain and also the region between the C _H 1 and C _h 2 domains, such that the interchain disulfide bond can be formed between the two heavy chains of two Fab 'fragments to form an F (ab') ₂ molecule.

An F (ab ') ₂ fragment contains two light chains and two heavy chains that contain a portion of the constant region between the C _H 1 and C _H 2 domains, such that an interchain disulfide bond is formed between the two heavy chains. In this way, an F (ab ') ₂ fragment is composed of two Fab' fragments that are held together by a disulfide bond between the two heavy chains.

The Fv region comprises the variable regions of both heavy and light chains, but is devoid of constant regions.

A domain antibody is an immunologically functional immunoglobulin fragment that contains only the variable region of a heavy chain or the variable region of a light chain. In some cases, two or more V _H regions are covalently joined with a peptide linker to create a divalent domain antibody. The two V _H regions of a divalent domain antibody can target the same antigen or different antigens.

A hemibody is an immunologically functional immunoglobulin construct that comprises a complete heavy chain, a complete light chain and a second heavy chain Fc region paired with the complete heavy chain Fc region. A linker can, but is not necessarily, employed to join the heavy chain Fc region and the second heavy chain Fc region. In particular embodiments, a hemibody is a monovalent form of an antigen-binding protein disclosed herein. In other modalities, pairs of loaded residues can be used to associate an Fc region with the second Fc region. The second heavy chain Fc region can comprise, for example, the ID. SEQ. N °: 441, and can be joined to the light chain by means of a linker (for example, SEQ ID. N °: 440). An exemplary hemibody heavy chain comprises the ID sequence. SEQ. No. 453.

A divalent antigen binding protein or divalent antibody comprises two antigen binding sites. In some cases, the two binding sites have the same antigen specificities. Bivalent antigen binding proteins and bivalent antibodies can be bispecific, as described herein.

A multispecific antigen or multispecific antibody binding protein is one that targets more than one antigen or epitope.

A bispecific, doubly specific or bifunctional antigen or antibody-binding protein is an antigen-binding protein or hybrid antibody, respectively, which has two different antigen-binding sites. Antigen-binding proteins and bispecific antibodies are a kind of multispecific antigen-binding protein or multispecific antibody and can be produced by a variety of methods, including, without limitation, fusion of hybridomas or binding of Fab 'fragments. See, for example, Songsivilai and Lachmann, 1990, Clin. Exp. Immunol. 79: 315-321; Kostelny et al, 1992, J. Immunol. 148: 1.547-1.553. The two binding sites of a bispecific antigen or antibody binding protein will bind to two different epitopes, which can reside on the same protein targets or on different protein targets.

The terms FGF21 type signaling and induces FGF21 type signaling, when applied to an antigen binding protein of the present disclosure, mean that the antigen binding protein mimics or modulates an in vivo biological effect induced by the binding of: (i) βKlotho; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 and induces a biological response that would otherwise result from binding FGF21 to: (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 in vivo. In assessing the binding and specificity of an antigen binding protein, for example, an antibody or immunologically functional fragment thereof, an antibody or fragment is considered to induce a biological response when the response is equal to or greater than 5% and, preferably, equal to or greater than 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80 %, 85%, 90% or 95% of the activity of a wild-type FGF21 pattern comprising the mature form of the ID. SEQ. N °: 2 (ie, the mature form of the human FGF21 sequence) and has the following properties: exhibits an efficiency level equal to or greater than 5% of a FGF21 standard, with an EC50 equal to or less than 100 nM, for example, 90 nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM or 10 nM: (1) in the recombinant cell-mediated luciferase reporter assay of Example 5 ; (2) ERK phosphorylation in the recombinant FGF21 receptor-mediated cell assay of Example 5; and (3) ERK phosphorylation in human adipocytes, as described in Example 7. The potency of an antigen binding protein is defined as exhibiting an EC50 equal to or less than 100 nM, for example, 90 nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM and preferably less than 10 nM of the antigen binding protein in the following assays: (1) the FGF21-mediated luciferase reporter cell assay recombinant from Example 5; (2) ERK phosphorylation in the recombinant FGF21 receptor-mediated cell assay of Example 5; and (3) phosphorylation of ERK in human adipocytes as described in Example 7.

It should be noted that not all antigen binding proteins of the present disclosure induce FGF21-mediated signaling, nor that this property is desirable in all circumstances. However, antigen binding proteins that do not induce FGF21-mediated signaling form aspects of the present disclosure and may be useful as diagnostic reagents or other applications.

As used herein, the term FGF21R means a multimeric receptor complex that FGF21 is known or supposed to form in vivo. In various embodiments, FGF21R comprises: (i) a FGFR, for example, FGFRlc, FGFR2c, FGFR3C or FGFR4, and (ii) β-Klotho.

The term polynucleotide or nucleic acid includes polymers of both single-stranded and double-stranded nucleotides. The nucleotides that comprise the polynucleotide can be ribonucleotides or deoxyribonucleotides or a modified form of one of the types of nucleotides. Said modifications include basic modifications such as, for example, bromouridine and inosine derivatives, modifications of ribose such as, for example, 2 ', 3'-dideoxyribose, and modifications of the internucleotide bond, such as, phosphorothiotate, phosphorodithiotate, phosphoroselenoate, phosphorodiselenoate, phosphoraniliotiotate, phosphoraniladate and phosphoramidate.

oligonucleotide means a polynucleotide comprising 200 or less nucleotides. In some modalities, oligonucleotides are 10 to 60 bases in length. In other embodiments, oligonucleotides are 12, 13, 14, 15, 16, 17, 18, 19, or 20 to 40 nucleotides in length. Oligonucleotides can be single-stranded or double-stranded, for example, for use in the construction of a mutant gene. Oligonucleotides can be sense or antisense oligonucleotides. An oligonucleotide can include a marker, including a radiolabel, a fluorescent marker, a hapten or an antigenic marker, for detection assays. Oligonucleotides can be used, for example, as PCR primers, cloning primers or hybridization probes.

An isolated nucleic acid molecule means DNA or RNA of genomic, mRNA, cDNA or synthetic origin or some combination thereof that is not associated with all or a portion of a polynucleotide in which the isolated polynucleotide is found in nature, or it is linked to a polynucleotide to which it is not linked in nature. For the purposes of this disclosure, it is understood that a nucleic acid molecule that comprises a particular nucleotide sequence does not comprise intact chromosomes. Isolated nucleic acid molecules that comprise specified nucleic acid sequences may include, in addition to the specified sequences, coding sequences for up to ten or even up to twenty other proteins or portions thereof, or may include operably linked regulatory sequences that control the expression of the region encoding the nucleic acid sequences cited and / or may include vector sequences.

Unless otherwise specified, the left end of any single strand polynucleotide sequence discussed here is the 5 'end; the direction to the left double stranded polynucleotide sequences is called the 5 'direction. The direction of adding 5 'to 3' of nascent RNA transcripts is termed as the direction of transcription;

regions of the sequence on the DNA strand that have the same sequence as the RNA transcript that are 5 'to the 5' end of the RNA transcript are called upstream sequences; regions of the sequence on the DNA strand that have the same sequence as the RNA transcript that are 3 'to the 3' end of the RNA transcript are called downstream sequences.

Control sequence refers to a sequence of polynucleotides that can affect the expression and processing of coding sequences to which it is linked. The nature of these control sequences may depend on the host organism. In particular embodiments, control sequences for prokaryotes can include a promoter, a ribosome binding site and a transcription termination sequence. For example, eukaryotic control sequences may include promoters that comprise one or several recognition sites for transcription factors, transcription enhancing sequences and transcription termination sequence. Control strings can include leader strings and / or merger partner strings.

vector means any molecule or entity (for example, nucleic acid, plasmid, bacteriophage or virus) used to transfer information from

coding in protein in a cell hostess. 0 term vector in expression or construction of expression if refers The a vector what is suitable for

transformation of a host cell and contains sequences of nucleic acids that direct and / or control (together with the host cell) the expression of one or more heterologous coding regions operatively linked to it. An expression construct can include, without limitation, sequences that affect or control transcription, translation and, if introns are present, affect the RNA splicing of a coding region operationally linked to it.

As used herein, the term operationally linked means that the components to which the term is applied are in a relationship that allows them to perform their inherent functions under suitable conditions. For example, a control sequence in a vector that is operationally linked to a protein coding sequence is linked to it in such a way that the expression of the protein coding sequence is obtained under conditions compatible with the transcriptional activity of the control sequences.

The term host cell means a cell that has been transformed, or is capable of being transformed, with a sequence of nucleic acids and thus expresses a gene of interest. The term includes the offspring of the parent cell, whether the offspring is identical or not in morphology or in genetic makeup to the original parent cell, as long as the gene of interest is present.

The term transduction means the transfer of genes from one bacterium to another, usually by bacteriophage. Transduction also refers to the acquisition and transfer of eukaryotic cell sequences by retrovirus with defective replication.

The term transfection means the uptake of foreign or exogenous DNA by a cell, and a cell was transfected when exogenous DNA was introduced into the cell's membrane. Various transfection methodologies are well known in the art and are disclosed herein. See, for example, Graham et al, (1973) Virology 52: 456; Sambrook et al., (2001) Molecular Cloning: A Laboratory Manual, supra; Davis et al., (1986) Basic Methods in Molecular Biology, Elsevier; Chu et al., (1981) Gene 13: 197. These techniques can be used to introduce one or more portions of exogenous DNA into suitable host cells.

The term transformation refers to a change in the genetic characteristics of a cell, and a cell was transformed when it was modified to contain new DNA or RNA. For example, a cell is transformed when it is genetically modified in relation to its native state by introducing new genetic material through transfection, transduction, or by other techniques. After transfection or transduction, the transforming DNA can recombine with that of the cell by physical integration into a cell chromosome, or it can be transiently maintained as an episomic element without being replicated, or it can replicate independently as a plasmid. A cell is considered to have been stably transformed when the transforming DNA is replicated by dividing the cell.

The terms polypeptide or protein are used interchangeably here to refer to a polymer of amino acid residues. The terms also apply to amino acid polymers in which one or more amino acid residues are an analogue or mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers. The terms also encompass polymers of amino acids that have been modified, for example, by adding carbohydrate residues to form glycoproteins, or phosphorylates. Polypeptides and proteins can be produced by a naturally occurring, non-recombinant cell, or polypeptides and proteins can be produced by a genetically modified or recombinant cell. Polypeptides and proteins can comprise molecules that have the amino acid sequence of a native protein, or molecules that have deletions, additions and / or substitutions for one or more amino acids in the native sequence. The terms polypeptide and protein encompass antigen-binding proteins that bind specifically or selectively to: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, or sequences that have deletions, additions and / or substitutions for one or more amino acids of an antigen-binding protein that specifically binds or selectively: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. The term polypeptide fragment refers to a polypeptide that has an amino terminal deletion, a carboxyl terminal deletion and / or an internal deletion, when compared to the full length protein. These fragments can also contain modified amino acids, when compared to the full-length protein. In certain embodiments, fragments are about five to 500 amino acids long. For example, fragments can be at least 5, 6, 8, 10,

14, 20, 50, 70, 100, 110, 150, 200, 250, 300, 350, 400 or 450 amino acids in length. Useful polypeptide fragments include immunologically functional fragments of antibodies, including binding domains. In the case of an antigen-binding protein that binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, useful fragments include, without limitation, a CDR region, a variable domain of a heavy or light chain, a portion of a chain antibody or just its variable region, including two CDRs, and the like.

The term isolated protein means that a protein in question: (1) is free of at least some other proteins with which it would normally be found, (2) is essentially free of other proteins from the same source, for example, from the same species, ( 3) it is expressed by a cell of a different species, (4) it has been separated from at least about 50 percent of polynucleotides, lipids, carbohydrates or other materials with which it is associated in nature, (5) it is operationally associated (by covalent or non-covalent interaction) with a polypeptide with which it is not associated in nature, or (6) does not occur in nature. Typically, an isolated protein constitutes at least about 5%, at least about 10%, at least about 25% or at least about 50% of a certain sample. Genomic DNA, cDNA, mRNA or other RNA, of synthetic origin, or any combination of these, can encode this isolated protein. Preferably, the isolated protein is substantially free of proteins or polypeptides or other contaminants that are found in its natural environment that would interfere with its therapeutic, diagnostic, prophylactic, research, or other use.

A variant of a polypeptide (for example, an antigen-binding protein, or an antibody) comprises an amino acid sequence in which one or more amino acid residues are inserted, deleted and / or replaced in the amino acid sequence in relation to another sequence polypeptide. Variants include fusion proteins.

A polypeptide derivative is a polypeptide (for example, an antigen binding protein, or an antibody) that has been chemically modified in some way other than insertion, deletion or substitution variants, for example, by conjugation with another chemical moiety.

naturally occurring term, as used throughout the specification in connection with biological materials such as, for example, polypeptides, nucleic acids, host cells, and the like, refers to materials that are found in nature.

The antigen binding region means a protein, or a portion of a protein, that specifically binds to a specified antigen, for example, FGFRlc, β-Klotho, or both FGFRlc and β-Klotho. For example, that portion of an antigen-binding protein that contains the amino acid residues that interact with an antigen and give the antigen-binding protein its specificity and affinity for the antigen is called an antigen-binding region. An antigen binding region typically includes one or more complementary binding regions (CDRs). Certain antigen binding regions also include one or more framework regions. A CDR is a sequence of amino acids that contributes to the specificity and affinity of antigen binding. Framework regions can assist in maintaining the proper conformation of CDRs to promote the link between the antigen binding region and an antigen.

In certain aspects, recombinant antigen binding proteins are provided that bind to (i) βKlotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. In this context, a recombinant protein is a protein made using recombinant techniques, that is, through the expression of a recombinant nucleic acid as described here. Methods and techniques for the production of recombinant proteins are well known in the art.

The term compete, when used in the context of antigen binding proteins (for example, neutralizing antigen binding proteins, neutralizing antibodies, agonist antigen binding proteins, agonist antibodies and binding proteins that bind to (i) β- Klotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4, which compete for the same epitope or binding site on a target, means competition between antigen binding proteins, as determined by an assay in which the antigen binding protein (eg, antibody or immunologically functional fragment thereof) under study prevents or inhibits specific binding of a reference molecule (eg, a ligand reference protein or reference antigen binding protein, for example, a reference antibody) to a common antigen (for example, FGFRlc, FGFR2c, FGFR3c, FGFR4, β-Klot ho, or a fragment thereof.) Several types of competitive binding assays can be used to determine whether a test molecule competes with a reference molecule for binding. Examples of assays that can be employed include direct or indirect solid phase radioimmunoassay (RIA), direct or indirect solid phase enzyme immunoassay (EIA), sandwich competition assay (see, for example, Stahli et al., (1983 ) Methods in Enzymology 9: 242-253); Direct EIA of solid phase biotinavidine (see, for example, Kirkland et al., (1986) J. Immunol. 137: 3.614-3.619); direct solid phase labeled assay, direct solid phase labeled sandwich assay (see, for example, Harlow and Lane, (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Press); Direct solid phase labeled RIA using 1-125 marker (see, for example, Morei et al., (1988) Molec. Immunol. 25: 7-15); Direct EIA of solid phase biotin-avidin (see, for example, Cheung, et al., (1990) Virology 176: 546-552); and direct labeled RIA (Moldenhauer et al., (1990) Scand. J. Immunol. 32: 77-82). Typically, such an assay involves the use of a purified antigen bound to a solid surface or cells that harbor an unlabeled test antigen binding protein or a marked reference antigen binding protein. Competitive inhibition is measured by determining the amount of marker attached to the solid surface or cells in the presence of the test antigen binding protein. Normally, the test antigen-binding protein is present in excess. Antigen-binding proteins identified by competition assay (competing antigen-binding proteins) include antigen-binding proteins to the same epitope as the reference antigen-binding proteins and antigen-binding proteins to an epitope adjacent sufficiently close to the epitope bound by the reference antigen binding protein for steric hindrance to occur. Additional details regarding methods for determining competitive binding will be provided in the examples presented here. Typically, when a competitive antigen binding protein is present in excess, it will exhibit specific binding of a reference antigen binding protein to a common antigen by at least 40%, 45%, 50%, 55%, 60%, 65%, 70% or 75%. In some cases, binding is inhibited by at least 80%, 85%, 90%, 95% or 97% or more.

The term antigen refers to a molecule or portion of a molecule capable of being bound by a selective binding agent, for example, an antigen binding protein (including, for example, an antibody or functional immune fragment thereof), and also it may be able to be used in an animal to produce antibodies capable of binding to that antigen. An antigen may have one or more epitopes that are capable of interacting with different antigen-binding proteins, for example, antibodies.

epitope means the amino acids of a target molecule that are brought into contact by an antigen binding protein (for example, an antibody) when the antigen binding protein is bound to the target molecule5. The term includes any subset of the complete list of amino acids in the target molecule that are brought into contact when an antigen-binding protein, for example, an antibody, is attached to the target molecule. An epitope can be contiguous or non-contiguous (for example, (i) 10 in a single-chain polypeptide, amino acid residues that are not contiguous with each other in the polypeptide sequence, but that within the context of the target molecule are linked by the antigen binding, or (ii) at a multimeric receptor comprising two or more 15 individual components, for example, (i) FGFRlc, FGFR2c,

FGFR3c or FGFR4, and (ii) β-Klotho, amino acid residues that are present in one or more of the individual components, but that are still linked by the antigen binding protein). In certain embodiments, epitopes may be mimetic, in that they comprise a three-dimensional structure that is similar to an antigenic epitope used to generate the antigen-binding protein, although they comprise none or only some of the amino acid residues found in that epitope used to generate the antigen-binding protein. Most often, epitopes reside in proteins, but in some cases, they can reside in other types of molecules, for example, nucleic acids. Epitope determinants can include chemically surfactant clusters of molecules such as, for example, amino acids, sugar side chains, phosphoryl or sulfonyl groups, and may have specific three-dimensional structural characteristics and / or specific charge characteristics. Generally, antigen-binding proteins specific to a particular target molecule will preferably recognize an epitope on the target molecule in a complex mixture of proteins and / or macromolecules.

The term identity refers to a relationship between the sequences of two or more polypeptide molecules or two or more nucleic acid molecules, as determined by sequence alignment and comparison. The term percent identity means the percentage of identical residues between amino acids or nucleotides in the compared molecules, and is calculated based on the size of the smallest of the molecules being compared. For these calculations, gaps (gaps) in alignments (if any) must be addressed by a particular mathematical model or computer program (ie, an algorithm). Methods that can be used to calculate the identity of aligned nucleic acids or polypeptides include those described in Computational Molecular Biology (Lesk, A.M., ed.) (1988) New York: Oxford University Press; Biocomputing Informatics and Genome Projects (Smith, D.W., ed.), 1993, New York: Academic Press; Computer Analysis of Sequence Data, Part I, (Griffin, A.M. and Griffin, H.G., eds.), 1994, New Jersey: Humana Press; von Heinje, G., (1987) Sequence Analysis in Molecular Biology, New York: Academic Press; Sequence Analysis Primer (Gribskov, M. and

Devereux, J., eds.), 1991, New York: M. Stockton Press; and Carillo et al., (1988) SIAM J. Applied Math. 48: 1,073.

In calculating the percentage identity, the sequences being compared are aligned in a way that generates the greatest combination between the sequences. The computer program used to determine percent identity is the GCG suite of programs, which includes GAP (Devereux et al., (1984) Nucl. Acid Res. 12: 387; Genetics Computer Group, University of Wisconsin, Madison, WI) . The GAP computer algorithm is used to align the two polypeptides or polynucleotides for which the percentage sequence identity is to be determined. The strings are aligned to optimally match their respective amino acids or nucleotides (the combined stretch, as determined by the algorithm). An open gap penalty (which is calculated as 3x the average diagonal, where the average diagonal is the mean of the diagonal of the comparison matrix being used; the diagonal is the score or number assigned to each perfect combination of amino acids by particular comparison matrix) and a gap extension penalty (which is usually 1/10 times the open gap penalty), as well as a comparison matrix such as PAM 250 or BLOSUM 62, are used in conjunction with the algorithm. In certain embodiments, a standard comparison matrix (see Dayhoff et al, (1978) Atlas of Protein Sequence and Structure 5: 345-352 for the PAM 250 comparison matrix; Henikoff et al, (1992) Proc. Natl Acad. Sci. USA 89: 10.915-10.919 for the BLOSUM 62 comparison matrix) is also used by the algorithm.

The recommended parameters for determining the percentage of identity for polypeptide or nucleotide sequences using the GAP program are as follows:

Algorithm: Needleman et al, 1970, J. Mol. Biol. 48: 443-453;

Comparison matrix: BLOSUM 62 by Henikoff et al, 1992, supra;

Gap penalty: 12 (but no penalty for end gaps);

Gap length penalty: 4;

Similarity threshold: 0.

Certain alignment schemes for aligning two sequences of amino acids can result in a combination of only a short region of the two sequences and that small aligned region may have very high sequence identity, although there is no significant relationship between the two full-length sequences. Consequently, the selected alignment method (for example, the GAP program) can be adjusted if so desired to result in an alignment that transposes at least 50 contiguous amino acids from the target polypeptide.

As used herein, substantially pure means that the species of molecule described is the predominant species present, that is, in molar terms it is more abundant than any other individual species in the same mixture. In certain embodiments, a substantially pure molecule is a composition in which the species in question comprises at least 50% (in molar terms) of all macromolecular species present. In other embodiments, a substantially pure composition will comprise at least 80%, 85%, 90%, 95% or 99% of all macromolecular species present in the composition. In other embodiments, the species in question is purified to the essential homogeneity in which contaminating species cannot be detected in the composition by conventional methods of detection and, therefore, the composition consists of a single detectable macromolecular species.

The terms treat and treatment refer to any evidence of success in treating or ameliorating an injury, pathology or condition, including any objective or subjective parameters such as, for example, withdrawal; remission; decrease in symptoms or make the injury, pathology or condition more tolerable for the patient; slow the rate of degeneration or decline; make the end point of degeneration less debilitating; improve the patient's physical or mental well-being. The treatment or improvement of symptoms can be based on objective or subjective parameters; including the results of a physical exam, neuropsychiatric exams and / or a psychiatric assessment. For example, certain methods presented here can be used to treat Type 2 diabetes, obesity and / or dyslipidemia, prophylactically or as an acute treatment, to lower blood glucose levels, to decrease circulating triglyceride levels, to decrease circulating levels of cholesterol and / or ameliorate a symptom associated with type 2 diabetes, obesity and dyslipidemia.

An effective amount is generally an amount sufficient to reduce the severity and / or frequency of symptoms, eliminate the symptoms and / or underlying cause, prevent the occurrence of symptoms and / or their underlying cause, and / or improve or remedy the damage that results or are associated with diabetes, obesity and dyslipidemia. In some embodiments, the effective amount is a therapeutically effective amount or a prophylactically effective amount. A therapeutically effective amount is an amount sufficient to remedy a disease state (for example, diabetes, obesity or dyslipidemia) or symptoms, particularly a condition or symptoms associated with the disease state, or otherwise prevent, prevent, delay or reverse the progression of the disease state or any other undesirable symptoms associated with the disease in any way. A prophylactically effective amount is an amount of a pharmaceutical composition that, when administered to an individual, will have the desired prophylactic effect, for example, preventing or delaying the onset (or recurrence) of diabetes, obesity or dyslipidemia, or reducing the likelihood of it (or recurrence) of diabetes, obesity or dyslipidemia or associated symptoms. The full therapeutic or prophylactic effect does not necessarily occur by administering a dose, and can occur only after administering a series of doses. Thus, a therapeutic or prophylactically effective amount can be administered in one or more administrations.

The term amino acid assumes its normal meaning in the art. The twenty naturally occurring amino acids and their abbreviations follow conventional usage. See Immunology-A Synthesis, 2nd Edition (ES Golub and DR Gren, eds.), Sinauer Associates: Sunderland, Mass. (1991), incorporated herein by reference for any purposes. Stereoisomers (eg, D-amino acids) of the twenty conventional amino acids, unnatural or unnaturally occurring amino acids, such as α, α-substituted amino acids, N-alkyl amino acids, and other unconventional amino acids, can also be suitable components for the polypeptides and are included in the phrase amino acid. Examples of non-naturally occurring amino acids (which can replace any naturally occurring amino acids found in any sequence disclosed herein, as desired) include: 4-hydroxyproline, γ-carboxyglutamate, ε-Ν, Ν, Ν-trimethillisine, ε-Ν-acetillisine, O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylisin, σ-Ν-methylylginine, and other similar amino acids and imino acids (for example, 4 hydroxyproline). In the polypeptide annotation used here, the left direction is the direction of the amino terminal and the right direction is the direction of the carboxyl terminal, according to standard usage and convention. A non-limiting list of examples of non-naturally occurring amino acids that can be inserted into an antigen-binding protein sequence or that can replace a wild-type residue in an antigen-binding sequence include βamino acids, homoamino acids, cyclic amino acids and amino acids with derivatized side chains. Examples include (in L or D form; abbreviated in parentheses): citrulline (Cit), homocitrulline (hCit), Ncxmethylcitrulina (NMeCit), Να-methylomocitrulin (NaMeHoCit), ornithine (Orn), Na-Methylornitine (Να-MeOrn or NMeOrn), sarcosine (Sar), homolysin (hLys or hK), homoarginine (hArg or hR), homoglutamine (hQ), Namethylarginine (NMeR), Na-metilleucine (Να-MeL or NMeL), Nmethylomolysin (NMeHoK), Να -methylglutamine (NMeQ), norleucine (Nle), norvaline (Nva), 1,2,3,4tetrahydroisoquinoline (Tic), octahidroindol-2-carboxylic acid (Oic), 3- (1-naphthyl) alanine (1-Nal), 3 - (2naftil) alanine (2-Nal), 1,2,3,4-tetrahydroisoquinoline (Tic), 2-indanylglycine (Igl), para-iodophenylalanine (plPhe), para-aminophenylalanine (4AmP or 4-Amino-Phe) , 4guanidino phenylalanine (Guf), glycillisine (abbreviated K (Νε-glycyl) or K (glycyl) or K (gly)), nitrophenylalanine (nitrophe), aminophenylalanine (aminophe or Amino-Phe), benzylphenylalanine (benzylphe), γ-carboxiglu), hydroxyproline (hydroxypro), p-carboxyl-phenylalanine (Cpa), αaminoadipic acid (Aad), Να-methyl valine (NMeVal), Na-methyl leucine (NMeLeu), Na-methylnorleucine (NMeNle), cyclopentylglycine (Cpg), cyclohexine Chg), acetylarginine (acetylarg), a, β-diaminopropionic acid (Dpr), α, γ-diaminobutyric acid (Dab), diaminopropionic acid (Dap), cyclohexylalanine (Cha), 4-methylphenylalanine (MePhe), β, β- diphenyl-alanine (BiPhA), aminobutyric acid (Abu), 4-phenyl-phenylalanine (or biphenylalanine; 4Bip), α-amino-isobutyric acid (Aib), beta-alanine, beta-aminopropionic acid, piperidinic acid, aminocaprioic acid, aminoheptanoic acid, aminopimelic acid, desmosine, diaminopimelic acid, Netylglycine, N-ethylparparin, hydroxylysine, hydroxylysine, hydroxylysine , allo-isoleucine, Nmethylglycine, N-methylisoleucine, N-methylvaline, 456 hydroxyproline (Hyp), γ-carboxyglutamate, ε-Ν, Ν, Νtrimetillisina, ε-Ν-acetillisine, 0-phosphoserine, N-methylethyl -methylhistidine, 5hydroxylysine, GJ-methylylginine, 4-amino-O-phthalic acid (4APA), and other similar amino acids, and derivative forms of any of those specifically listed.

II. OVERVIEW

Binding proteins that bind to (i) β-Klotho are provided herein; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. A unique property of the antigen binding proteins disclosed here is the agonist nature of these proteins, specifically the ability to mimic the in vivo effect of FGF21 and to induce FGF21-like signaling. More specifically strikingly, some of the antigen binding proteins disclosed herein induce FGF21-like signaling in various in vitro cell-based assays, including the ELK-luciferase reporter from Example 5 under the following conditions: (1) binding and FGF21 receptor activity is β-Klotho-dependent; (2) the activity is selective for FGFRlc / β-Klotho complex; (3) binding to FGFRlc / βKlotho triggers FGF21-like signaling pathways; and (4) the potency (EC50) is comparable to a wild-type FGF21 standard comprising the mature form of the ID. SEQ. No. 2 as measured in the following cell-based assays: (1) the recombinant FGF21 receptor-mediated luciferase reporter assay of Example 5;

(2) ERK phosphorylation in the recombinant FGF21 receptor-mediated cell assay of Example 5; and (3) ERK phosphorylation in human adipocytes, as described in more detail in Example 7. The disclosed antigen binding proteins are therefore expected to exhibit in vivo activities that are consistent with the natural biological function of FGF21. This property makes the antigen binding proteins revealed to be viable therapeutic substances for the treatment of metabolic diseases such as, for example, type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease, metabolic syndrome and widely any disease or condition in which it is desirable to mimic or increase the in vivo effects of FGF21.

In some embodiments of the present disclosure, the provided antigen-binding proteins may comprise polypeptides in which one or more complementarity determining regions (CDRs) may be embedded and / or joined. In these antigen binding proteins, the CDRs can be embedded in a framework region, which guides the CDR (s) in such a way that the appropriate antigen binding properties of the CDR (s) are obtained. In general, these antigen binding proteins that are provided can facilitate or enhance the interaction between FGFRlc and β-Klotho, and can substantially induce FGF21-like signaling.

Certain antigen binding proteins described herein are antibodies or are derived from antibodies. In certain embodiments, the polypeptide structure of antigen binding proteins is based on antibodies, including, without limitation, monoclonal antibodies, bispecific antibodies, minibodies, domain antibodies, synthetic antibodies (sometimes referred to as antibody mimetics), chimeric antibodies, humanized antibodies, human antibodies, antibody fusions (sometimes referred to herein as antibody conjugates), hemibodies, and fragments thereof. The various structures will be further described here below.

The antigen binding proteins provided herein have been shown to bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4, and particularly to: (i) human β-Klotho; (ii) human FGFRlc, human FGFR2c, human FGFR3c or human FGFR4; or (iii) a complex comprising human β-Klotho and one of human FGFRlc, human FGFR2c, human FGFR3c and human FGFR4. As described and shown in the Examples presented here, based on Western blot results, commercially available anti-βKlotho or anti-FGFRlc antibodies bind to denatured β-Klotho or FGFRlc, while the antigen binding protein (agonist antibodies) does not does. Conversely, the provided antigen binding proteins recognize the native structure of FGFRlc and βKlotho on the cell surface, whereas commercial antibodies do not, based on the FACS results provided. See Example 9. The antigen binding proteins that are provided, therefore, mimic the natural biological activity of FGF21 in vivo. As a result, the antigen binding proteins provided here are capable of activating FGF21-like signaling activity. In particular, the revealed antigen-binding proteins may have one or more of the following in vivo activities: induction of FGF21-like signal transduction pathways, reduced blood glucose levels, reduced circulating lipid levels, improved metabolic parameters and other physiological effects induced in vivo by the formation of the ternary complex of FGFRlc, β-Klotho and FGF21, for example, in conditions such as, for example, type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease and metabolic syndrome.

Antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 which are disclosed herein have a variety of uses. Some of the antigen binding proteins, for example, are useful in specific binding assays, in the affinity purification of: (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, including the human forms of these revealed proteins, and in evaluation assays to identify other agonists of the signaling activity of the

type FGF21. antigen what if call Proteins in Link specifically to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) one complex which comprises β-Klotho and a

of FGFRlc, FGFR2c, FGFR3c and FGFR4 which are disclosed herein can be used in various treatment applications, as explained here. For example, certain antigen-binding proteins are useful for treating conditions associated with FGF21-like signaling processes in a patient, for example, reduction, relief or treatment of type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease and metabolic syndrome. Other uses for antigen binding proteins include, for example, diagnosis of diseases or conditions associated with β-Klotho, FGFRlc, FGFR2C, FGFR3c, FGFR4 or FGF21, and evaluation assays to determine the presence or absence of these molecules. Some of the antigen binding proteins described herein may be useful in the treatment of conditions, symptoms and / or the pathology associated with decreased FGF21-like signaling activity. Exemplary conditions include, without limitation, diabetes, obesity, NASH and dyslipidemia.

FGF21

The antigen binding proteins disclosed herein induce FGF21-mediated signaling, as defined herein. In vivo, the mature form of FGF21 is the active form of the molecule. The nucleotide sequence encoding full length FGF21 is provided; the nucleotides encoding the signal sequence are underlined.

ATG GAC TCG GAC GAG ACC GGG TTC GAG CAC TCA GGA CTG TGG GTT TCT GTG CTG GCT GGT CTT CTG CTG GGA GCC TGC CAG GCA CAC CCC ATC CCT GAC CBT AGT CCT CTC CTG CAA TTC GGG GGC CAA GTC CGG CAG CGG TAC CTC TAC A CA GAT GAT GCC CAG CAG A CA GAA GCC CAC CTG GAG ATC AGG GAG GAT GGG ACG GTG GGG GGC GCT GCT GAC CAG AGC CCC GAA AGT CTC CTG CAG CTG AAA GCC TTG THE AG CCG GGA GTT ATT CAA ATC TTG GGA GTC THE AG A CA CBT AGG TTC CTG TGC CAG CGG CCA GAT GGG GCC CTG TAT GGA TCG CTC CAC TTT GAC CCT GAG GCC TGC AGC TTC CGG GAG CTG CTT CTT GAG GAC GGA TAC AAT GTT TAC CAG CBT GAA GCC CAC GGC CTC CCG CTG CAC CTG CCA GGG AAC THE AG

CBT CCA CAC CGG GAC CCT GCA CCC CGA GGA CCA GCT CGC TTC CTG CCA CTA CCA GGC CTG CCC CCC GCA CCC CCG GAG CCA CCC GGA ATC CTG GCC CCC CAG CCC CCC GAT GTG GGC CBT TCG GAC CCT CTG AGC ATG GTG GGA CCT CBT CAG GGC CGA AGC CCC AGC TAC GCT CBT TGA (ID. . IN SEQ. . N °: : D.

The full length FGF21 amino acid sequence is provided; amino acids constituting the sequence are underlined Signal: S MDSDETGFEHSGLWVSVLAGLLLGACQAHPIPDS PLLQFGGQVRQRYLYTDDAQQTEA HLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDP EACSFRELLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPAPPEP PGILAPQPPDVGSSDPLSMVGPSQGRSPSYAS (SEQ ID NO:.. 2).

FGFRlc

The antigen binding proteins disclosed herein bind FGFRlc, in particular human FGFRlc, when associated with β-Klotho. The nucleotide sequence encoding human FGFRlc Number (GenBank Accession No. NM_023110) is provided: ATGTGGAGCTGGAAGTGCCTCCTCTTCTGGGCTGTGCTGGTCACAGCCACACTCTGCAC CGCTAGGCCGTCCCCGACCTTGCCTGAACAAGCCCAGCCCTGGGGAGCCCCTGTGGAAG TGGAGTCCTTCCTGGTCCACCCCGGTGACCTGCTGCAGCTTCGCTGTCGGCTGCGGGAC GATGTGCAGAGCATCAACTGGCTGCGGGACGGGGTGCAGCTGGCGGAAAGCAACCGCAC CCGCATCACAGGGGAGGAGGTGGAGGTGCAGGACTCCGTGCCCGCAGACTCCGGCCTCT ATGCTTGCGTAACCAGCAGCCCCTCGGGCAGTGACACCACCTACTTCTCCGTCAATGTT TCAGATGCTCTCCCCTCCTCGGAGGATGATGATGATGATGATGACTCCTCTTCAGAGGA GAAAGAAACAGATAACACCAAACCAAACCGTATGCCCGTAGCTCCATATTGGACATCAC CAGAAAAGATGGAAAAGAAATTGCATGCAGTGCCGGCTGCCAAGACAGTGAAGTTCAAA TGCCCTTCCAGTGGGACACCAAACCCAACACTGCGCTGGTTGAAAAATGGCAAAGAATT CAAACCTGACCACAGAATTGGAGGCTACAAGGTCCGTTATGCCACCTGGAGCATCATAA TGGACTCTGTGGTGCCCTCTGACAAGGGCAACTACACCTGCATTGTGGAGAATGAGTAC

GGCAGCATCAACCACACATACCAGCTGGATGTCGTGGAGCGGTCCCCTCACCGGCCCAT CCTGCAAGCAGGGTTGCCCGCCAACAAAACAGTGGCCCTGGGTAGCAACGTGGAGTTCA TGTGTAAGGTGTACAGTGACCCGCAGCCGCACATCCAGTGGCTAAAGCACATCGAGGTG AATGGGAGCAAGATTGGCCCAGACAACCTGCCTTATGTCCAGATCTTGAAGACTGCTGG AGTTAATACCACCGACAAAGAGATGGAGGTGCTTCACTTAAGAAATGTCTCCTTTGAGG ACGCAGGGGAGTATACGTGCTTGGCGGGTAACTCTATCGGACTCTCCCATCACTCTGCA TGGTTGACCGTTCTGGAAGCCCTGGAAGAGAGGCCGGCAGTGATGACCTCGCCCCTGTA CCTGGAGATCATCATCTATTGCACAGGGGCCTTCCTCATCTCCTGCATGGTGGGGTCGG TCATCGTCTACAAGATGAAGAGTGGTACCAAGAAGAGTGACTTCCACAGCCAGATGGCT GTGCACAAGCTGGCCAAGAGCATCCCTCTGCGCAGACAGGTAACAGTGTCTGCTGACTC CAGTGCATCCATGAACTCTGGGGTTCTTCTGGTTCGGCCATCACGGCTCTCCTCCAGTG GGACTCCCATGCTAGCAGGGGTCTCTGAGTATGAGCTTCCCGAAGACCCTCGCTGGGAG CTGCCTCGGGACAGACTGGTCTTAGGCAAACCCCTGGGAGAGGGCTGCTTTGGGCAGGT GGTGTTGGCAGAGGCTATCGGGCTGGACAAGGACAAACCCAACCGTGTGACCAAAGTGG CTGTGAAGATGTTGAAGTCGGACGCAACAGAGAAAGACTTGTCAGACCTGATCTCAGAA ATGGAGATGATGAAGATGATCGGGAAGCATAAGAATATCATCAACCTGCTGGGGGCCTG CACGCAGGATGGTCCCTTGTATGTCATCGTGGAGTATGCC TCCAAGGGCAACCTGCGGG AGTACCTGCAGGCCCGGAGGCCCCCAGGGCTGGAATACTGCTACAACCCCAGCCACAAC CCAGAGGAGCAGCTCTCCTCCAAGGACCTGGTGTCCTGCGCCTACCAGGTGGCCCGAGG CATGGAGTATCTGGCCTCCAAGAAGTGCATACACCGAGACCTGGCAGCCAGGAATGTCC TGGTGACAGAGGACAATGTGATGAAGATAGCAGACTTTGGCCTCGCACGGGACATTCAC CACATCGACTACTATAAAAAGACAACCAACGGCCGACTGCCTGTGAAGTGGATGGCACC CGAGGCATTATTTGACCGGATCTACACCCACCAGAGTGATGTGTGGTCTTTCGGGGTGC TCCTGTGGGAGATCTTCACTCTGGGCGGCTCCCCATACCCCGGTGTGCCTGTGGAGGAA CTTTTCAAGCTGCTGAAGGAGGGTCACCGCATGGACAAGCCCAGTAACTGCACCAACGA GCTGTACATGATGATGCGGGACTGCTGGCATGCAGTGCCCTCACAGAGACCCACCTTCA AGCAGCTGGTGGAAGACCTGGACCGCATCGTGGCCTTGACCTCCAACCAGGAGTACCTG GACCTGTCCATGCCCCTGGACCAGTACTCCCCCAGCTTTCCCGACACCCGGAGCTCTAC GTGCTCCTCAGGGGAGGATTCCGTCTTCTCTCATGAGCCGCTGCCCGAGGAGCCCTGCC

TGCCCCGACACCCAGCCCAGCTTGCCAATGGCGGACTCAAACGCCGCTGA (SEQ ID. NO: 3).

The amino acid sequence of human FGFRlc (GenBank Accession Number NP_075598) is provided:

MWSWKCLLFWAVLVTATLCTARPSPTLPEQAQPWGAPVEVESFLVHPGDLLQLRCRLRD DVQSINWLRDGVQLAESNRTRITGEEVEVQDSVPADSGLYACVTSSPSGSDTTYFSVNV SDALPSSEDDDDDDDSSSEEKETDNTKPNRMPVAPYWTSPEKMEKKLHAVPAAKTVKFK CPSSGTPNPTLRWLKNGKEFKPDHRIGGYKVRYATWSIIMDSWPSDKGNYTCIVENEY GSINHTYQLDWERSPHRPILQAGLPANKTVALGSNVE FMCKVYSDPQPHIQWLKHIEV NGSKIGPDNLPYVQILKTAGVNTTDKEMEVLHLRNVSFEDAGEYTCLAGNSIGLSHHSA WLTVLEALEERPAVMTSPLYLEIIIYCTGAFLISCMVGSVIVYKMKSGTKKSDFHSQMA VHKLAKSIPLRRQVTVSADSSASMNSGVLLVRPSRLSSSGTPMLAGVSEYELPEDPRWE LPRDRLVLGKPLGEGCFGQWLAEAIGLDKDKPNRVTKVAVKMLKSDATEKDLSDLISE MEMMKMIGKHKNIINLLGACTQDGPLYVIVEYASKGNLREYLQARRPPGLEYCYNPSHN PEEQLSSKDLVSCAYQVARGMEYLASKKCIHRDLAARNVLVTEDNVMKIADFGLARDIH HIDYYKKTTNGRLPVKWMAPEALFDRIYTHQSDVWSFGVLLWEIFTLGGSPYPGVPVEE LFKLLKEGHRMDKPSNCTNELYMMMRDCWHAVPSQRPTFKQLVEDLDRIVALTSNQEYL DLSMPLDQYSPSFPDTRSSTCSSGEDSVFSHEPLPEEPCLPRHPAQLANGGLKRR (SEQ ID NO:.. 4).

The antigen binding proteins described herein bind to the extracellular portion of FGFRlc. An example of an FGFRlc extracellular region is:

MWSWKCLLFWAVLVTATLCTARPSPTLPEQAQPWGAPVEVESFLVHPGDLLQLRCRLRD DVQSINWLRDGVQLAESNRTRITGEEVEVQDSVPADSGLYACVTSSPSGSDTTYFSVNV SDALPSSEDDDDDDDSSSEEKETDNTKPNRMPVAPYWTSPEKMEKKLHAVPAAKTVKFK CPSSGTPNPTLRWLKNGKEFKPDHRIGGYKVRYATWSIIMDSWPSDKGNYTCIVENEY GSINHTYQLDWERSPHRPILQAGLPANKTVALGSNVEFMCKVYSDPQPHIQWLKHIEV NGSKIGPDNLPYVQILKTAGVNTTDKEMEVLHLRNVSFEDAGEYTCLAGNSIGLSHHSA WLTVLEALEERPAVMTSPLY (SEQ ID NO:.. 5).

As described herein, FGFR1c proteins can also include fragments. As used herein, the terms are used interchangeably to mean a receptor, in particular, and, unless otherwise specified, a human receptor that, in association with β-Klotho and FGF21, induces FGF21-like signaling activity .

FGFRlc term also includes post-translational modifications of the FGFRlc amino acid sequence, for example, possible glycosylation sites linked to N. Thus, antigen binding proteins can bind or be generated by glycosylated proteins at one or more of the positions .

β-Klotho

The antigen binding proteins disclosed herein bind to β-Klotho, in particular human β-Klotho. The nucleotide sequence encoding human β-Klotho (GenBank Accession Number NM_175737) is provided:

ATGAAGCCAGGCTGTGCGGCAGGATCTCCAGGGAATGAATGGATTTTCTTCAGCACTGA TGAAATAACCACACGCTATAGGAATACAATGTCCAACGGGGGATTGCAAAGATCTGTCA TCCTGTCAGCACTTATTCTGCTACGAGCTGTTACTGGATTCTCTGGAGATGGAAGAGCT ATATGGTCTAAAAATCCTAATTTTACTCCGGTAAATGAAAGTCAGCTGTTTCTCTATGA CACTTTCCCTAAAAACTTTTTCTGGGGTATTGGGACTGGAGCATTGCAAGTGGAAGGGA GTTGGAAGAAGGATGGAAAAGGACCTTCTATATGGGATCATTTCATCCACACACACCTT AAAAATGTCAGCAGCACGAATGGTTCCAGTGACAGTTATATTTTTCTGGAAAAAGACTT ATCAGCCCTGGATTTTATAGGAGTTTCTTTTTATCAATTTTCAATTTCCTGGCCAAGGC TTTTCCCCGATGGAATAGTAACAGTTGCCAACGCAAAAGGTCTGCAGTACTACAGTACT CTTCTGGACGCTCTAGTGCTTAGAAACATTGAACCTATAGTTACTTTATACCACTGGGA TTTGCCTTTGGCACTACAAGAAAAATATGGGGGGTGGAAAAATGATACCATAATAGATA TCTTCAATGACTATGCCACATACTGTTTCCAGATGTTTGGGGACCGTGTCAAATATTGG ATTACAATTCACAACCCATATCTAGTGGCTTGGCATGGGTATGGGACAGGTATGCATGC

CCCTGGAGAGAAGGGAAATTTAGCAGCTGTCTACACTGTGGGACACAACTTGATCAAGG

CTCACTCGAAAGTTTGGCATAACTACAACACACATTTCCGCCCACATCAGAAGGGTTGG

TTATCGATCACGTTGGGATCTCATTGGATCGAGCCAAACCGGTCGGAAAACACGATGGA

TATATTCAAATGTCAACAATCCATGGTTTCTGTGCTTGGATGGTTTGCCAACCCTATCC

ATGGGGATGGCGACTATCCAGAGGGGATGAGAAAGAAGTTGTTCTCCGTTCTACCCATT

TTCTCTGAAGCAGAGAAGCATGAGATGAGAGGCACAGCTGATTTCTTTGCCTTTTCTTT

TGGACCCAACAACTTCAAGCCCCTAAACACCATGGCTAAAATGGGACAAAATGTTTCAC

TTAATTTAAGAGAAGCGCTGAACTGGATTAAACTGGAATACAACAACCCTCGAATCTTG

ATTGCTGAGAATGGCTGGTTCACAGACAGTCGTGTGAAAACAGAAGACACCACGGCCAT

CTACATGATGAAGAATTTCCTCAGCCAGGTGCTTCAAGCAATAAGGTTAGATGAAATAC

GAGTGTTTGGTTATACTGCCTGGTCTCTCCTGGATGGCTTTGAATGGCAGGATGCTTAC

ACCATCCGCCGAGGATTATTTTATGTGGATTTTAACAGTAAACAGAAAGAGCGGAAACC

TAAGTCTTCAGCACACTACTACAAACAGATCATACGAGAAAATGGTTTTTCTTTAAAAG

AGTCCACGCCAGATGTGCAGGGCCAGTTTCCCTGTGACTTCTCCTGGGGTGTCACTGAA

TCTGTTCTTAAGCCCGAGTCTGTGGCTTCGTCCCCACAGTTCAGCGATCCTCATCTGTA

CGTGTGGAACGCCACTGGCAACAGACTGTTGCACCGAGTGGAAGGGGTGAGGCTGAAAA

CACGACCCGCTCAATGCACAGATTTTGTAAACATCAAAAAACAACTTGAGATGTTGGCA

AGAATGAAAGTCACCCACTACCGGTTTGCTCTGGATTGGGCCTCGGTCCTTCCCACTGG

CAACCTGTCCGCGGTGAACCGACAGGCCCTGAGGTACTACAGGTGCGTGGTCAGTGAGG

GGCTGAAGCTTGGCATCTCCGCGATGGTCACCCTGTATTATCCGACCCACGCCCACCTA

GGCCTCCCCGAGCCTCTGTTGCATGCCGACGGGTGGCTGAACCCATCGACGGCCGAGGC

CTTCCAGGCCTACGCTGGGCTGTGCTTCCAGGAGCTGGGGGACCTGGTGAAGCTCTGGA

TCACCATCAACGAGCCTAACCGGCTAAGTGACATCTACAACCGCTCTGGCAACGACACC

TACGGGGCGGCGCACAACCTGCTGGTGGCCCACGCCCTGGCCTGGCGCCTCTACGACCG

GCAGTTCAGGCCCTCACAGCGCGGGGCCGTGTCGCTGTCGCTGCACGCGGACTGGGCGG

AACCCGCCAACCCCTATGCTGACTCGCACTGGAGGGCGGCCGAGCGCTTCCTGCAGTTC

GAGATCGCCTGGTTCGCCGAGCCGCTCTTCAAGACCGGGGACTACCCCGCGGCCATGAG

GGAATACATTGCCTCCAAGCACCGACGGGGGCTTTCCAGCTCGGCCCTGCCGCGCCTCA

CCGAGGCCGAAAGGAGGCTGCTCAAGGGCACGGTCGACTTCTGCGCGCTCAACCACTTC

ACCACTAGGTTCGTGATGCACGAGCAGCTGGCCGGCAGCCGCTACGACTCGGACAGGGA

CATCCAGTTTCTGCAGGACATCACCCGCCTGAGCTCCCCCACGCGCCTGGCTGTGATTC CCTGGGGGGTGCGCAAGCTGCTGCGGTGGGTCCGGAGGAACTACGGCGACATGGACATT TACATCACCGCCAGTGGCATCGACGACCAGGCTCTGGAGGATGACCGGCTCCGGAAGTA CTACCTAGGGAAGTACCTTCAGGAGGTGCTGAAAGCATACCTGATTGATAAAGTCAGAA TCAAAGGCTATTATGCATTCAAACTGGCTGAAGAGAAATCTAAACCCAGATTTGGATTC TTCACATCTGATTTTAAAGCTAAATCCTCAATACAATTTTACAACAAAGTGATCAGCAG CAGGGGCTTCCCTTTTGAGAACAGTAGTTCTAGATGCAGTCAGACCCAAGAAAATACAG AGTGCACTGTCTGCTTATTCCTTGTGCAGAAGAAACCACTGATATTCCTGGGTTGTTGC TTCTTCTCCACCCTGGTTCTACTCTTATCAATTGCCATTTTTCAAAGGCAGAAGAGAAG AAAGTTTTGGAAAGCAAAAAACTTACAACACATACCATTAAAGAAAGGCAAGAGAGTTG TTAGCTAA (SEQ ID NO:.. 6).

The sequence of human β-Klotho amino acids in total length (number of GenBank Accession No. NP_783864) is provided: MKPGCAAGSPGNEWIFFSTDEITTRYRNTMSNGGLQRSVILSALILLRAVTGFSGDGRA IWSKNPNFTPVNESQLFLYDTFPKNFFWGIGTGALQVEGSWKKDGKGPSIWDHFIHTHL KNVSSTNGSSDSYIFLEKDLSALDFIGVSFYQFSISWPRLFPDGIVTVANAKGLQYYST LLDALVLRNIEPIVTLYHWDLPLALQEKYGGWKNDTIIDIFNDYATYCFQMFGDRVKYW ITIHNPYLVAWHGYGTGMHAPGEKGNLAAVYTVGHNLIKAHSKVWHNYNTHFRPHQKGW LSITLGSHWIEPNRS ENTMDIFKCQQSMVSVLGWFANPIHGDGDYPEGMRKKL FSVL PI FS EAE KHEMRGTAD F FAFS FGPNNFKPLNTMAKMGQNVS LNLREALNWIKLEYNNPRIL IAENGWFTDSRVKTEDTTAIYMMKNFLSQVLQAIRLDEIRVFGYTAWSLLDGFEWQDAY TIRRGLFYVDFNSKQKERKPKSSAHYYKQIIRENGFSLKESTPDVQGQFPCDFSWGVTE SVLKPESVASSPQFSDPHLYVWNATGNRLLHRVEGVRLKTRPAQCTDFVNIKKQLEMLA RMKVTHYRFALDWASVLPTGNLSAVNRQALRYYRCVVSEGLKLGISAMVTLYYPTHAHL GLPEPLLHADGWLNPSTAEAFQAYAGLCFQELGDLVKLWITINEPNRLSDIYNRSGNDT YGAAHNLLVAHALAWRLYDRQFRPSQRGAVSLSLHADWAEPANPYADSHWRAAERFLQF EIAWFAEPLFKTGDYPAAMREYIASKHRRGLSSSALPRLTEAERRLLKGTVDFCALNHF TTRFVMHEQLAGSRYDSDRDIQFL QDITRLSSPTRLAVIPWGVRKLLRWVRRNYGDMDI YITASGIDDQALEDDRLRKYYLGKYLQEVLKAYLIDKVRIKGYYAFKLAEEKSKPRFGF

FTSDFKAKSSIQFYNKVISSRGFPFENSSSRCSQTQENTECTVCLFLVQKKPLIFLGCC FFSTLVLLLSIAIFQRQKRRKFWKAKNLQHIPLKKGKRWS (SEQ ID: 7).

The antigen binding proteins described here bind to the extracellular portion of β-Klotho. An example of an extracellular region of β-Klotho is: MKPGCAAGSPGNEWIFFSTDEITTRYRNTMSNGGLQRSVILSALILLRAVTGFSGDGRA IWSKNPNFTPVNESQLFLYDTFPKNFFWGIGTGALQVEGSWKKDGKGPSIWDHFIHTHL KNVSSTNGSSDSYIFLEKDLSALDFIGVSFYQFSISWPRLFPDGIVTVANAKGLQYYST LLDALVLRNIEPIVTLYHWDLPLALQEKYGGWKNDTIIDIFNDYATYCFQMFGDRVKYW ITIHNPYLVAWHGYGTGMHAPGEKGNLAAVYTVGHNLIKAHS KVWHNYNTHFRPHQKGW LSITLGSHWIEPNRSENTMDIFKCQQSMVSVLGWFANPIHGDGDYPEGMRKKLFSVLPI FSEAEKHEMRGTADFFAFSFGPNNFKPLNTMAKMGQNVSLNLREALNWIKLEYNNPRIL IAENGWFTDSRVKTEDTTAIYMMKNFLSQVLQAIRLDEIRVFGYTAWSLLDGFEWQDAY TIRRGLFYVDFNSKQKERKPKSSAHYYKQIIRENGFSLKESTPDVQGQFPCDFSWGVTE SVLKPESVASSPQFSDPHLYVWNATGNRLLHRVEGVRLKTRPAQCTDFVNIKKQLEMLA RMKVTHYRFALDWASVLPTGNLSAVNRQALRYYRCVVSEGLKLGISAMVTLYYPTHAHL GLPEPLLHADGWLNPSTAEAFQAYAGLCFQELGDLVKLWITINEPNRLSDIYNRSGNDT YGAAHNLLVAHALAWRLYDRQFRPSQRGAVSLSLHADWAEPANPYADSHWRAAERFLQF EIAWFAEPLFKTGDYPAAMREYIASKHRRGLSSSALPRLTEAERRLLKGTVDFCALNHF TTRFVMHEQLAGSRYDSDRDIQFLQDITRLSSPTRLAVIPWGVRKLLRWVRRNYGDMDI YITASGIDDQALEDDRLRKYYLGKYLQEVLKAYLIDKVRIKGY YAFKLAEEKSKPRFGF FTSDFKAKSSIQFYNKVISSRGFPFENSSSRCSQTQENTECTVCLFLVQKKP (ID. SEQ. No. 8).

The murine form of β-Klotho, and fragments and subsequences thereof, may be useful in the study and / or construction of the molecules provided here. The nucleotide sequence encoding murine β-Klotho (GenBank Accession Number NM_031180) is provided:

ATGAAGACAGGCTGTGCAGCAGGGTCTCCGGGGAATGAATGGATTTTCTTCAGCTCTGA

TGAAAGAAACACACGCTCTAGGAAAACAATGTCCAACAGGGCACTGCAAAGATCTGCCG

TGCTGTCTGCGTTTGTTCTGCTGCGAGCTGTTACCGGCTTCTCCGGAGACGGGAAAGCA

ATATGGGATAAAAAACAGTACGTGAGTCCGGTAAACCCAAGTCAGCTGTTCCTCTATGA

CACTTTCCCTAAAAACTTTTCCTGGGGCGTTGGGACCGGAGCATTTCAAGTGGAAGGGA

GTTGGAAGACAGATGGAAGAGGACCCTCGATCTGGGATCGGTACGTCTACTCACACCTG

AGAGGTGTCAACGGCACAGACAGATCCACTGACAGTTACATCTTTCTGGAAAAAGACTT

GTTGGCTCTGGATTTTTTAGGAGTTTCTTTTTATCAGTTCTCAATCTCCTGGCCACGGT

TGTTTCCCAATGGAACAGTAGCAGCAGTGAATGCGCAAGGTCTCCGGTACTACCGTGCA

CTTCTGGACTCGCTGGTACTTAGGAATATCGAGCCCATTGTTACCTTGTACCATTGGGA

TTTGCCTCTGACGCTCCAGGAAGAATATGGGGGCTGGAAAAATGCAACTATGATAGATC

TCTTCAACGACTATGCCACATACTGCTTCCAGACCTTTGGAGACCGTGTCAAATATTGG

ATTACAATTCACAACCCTTACCTTGTTGCTTGGCATGGGTTTGGCACAGGTATGCATGC

ACCAGGAGAGAAGGGAAATTTAACAGCTGTCTACACTGTGGGACACAACCTGATCAAGG

CACATTCGAAAGTGTGGCATAACTACGACAAAAACTTCCGCCCTCATCAGAAGGGTTGG

CTCTCCATCACCTTGGGGTCCCATTGGATAGAGCCAAACAGAACAGACAACATGGAGGA

CGTGATCAACTGCCAGCACTCCATGTCCTCTGTGCTTGGATGGTTCGCCAACCCCATCC

ACGGGGACGGCGACTACCCTGAGTTCATGAAGACGGGCGCCATGATCCCCGAGTTCTCT

GAGGCAGAGAAGGAGGAGGTGAGGGGCACGGCTGATTTCTTTGCCTTTTCCTTCGGGCC

CAACAACTTCAGGCCCTCAAACACCGTGGTGAAAATGGGACAAAATGTATCACTCAACT

TAAGGCAGGTGCTGAACTGGATTAAACTGGAATACGATGACCCTCAAATCTTGATTTCG

GAGAACGGCTGGTTCACAGATAGCTATATAAAGACAGAGGACACCACGGCCATCTACAT

GATGAAGAATTTCCTAAACCAGGTTCTTCAAGCAATAAAATTTGATGAAATCCGCGTGT

TTGGTTATACGGCCTGGACTCTCCTGGATGGCTTTGAGTGGCAGGATGCCTATACGACC

CGACGAGGGCTGTTTTATGTGGACTTTAACAGTGAGCAGAAAGAGAGGAAACCCAAGTC

CTCGGCTCATTACTACAAGCAGATCATACAAGACAACGGCTTCCCTTTGAAAGAGTCCA

CGCCAGACATGAAGGGTCGGTTCCCCTGTGATTTCTCTTGGGGAGTCACTGAGTCTGTT

CTTAAGCCCGAGTTTACGGTCTCCTCCCCGCAGTTTACCGATCCTCACCTGTATGTGTG

GAATGTCACTGGCAACAGATTGCTCTACCGAGTGGAAGGGGTAAGGCTGAAAACAAGAC

CATCCCAGTGCACAGATTATGTGAGCATCAAAAAACGAGTTGAAATGTTGGCAAAAATG

AAAGTCACCCACTACCAGTTTGCTCTGGACTGGACCTCTATCCTTCCCACTGGCAATCT

GTCCAAAGTTAACAGACAAGTGTTAAGGTACTATAGGTGTGTGGTGAGCGAAGGACTGA AGCTGGGCGTCTTCCCCATGGTGACGTTGTACCACCCAACCCACTCCCATCTCGGCCTC CCCCTGCCACTTCTGAGCAGTGGGGGGTGGCTAAACATGAACACAGCCAAGGCCTTCCA GGACTACGCTGAGCTGTGCTTCCGGGAGTTGGGGGACTTGGTGAAGCTCTGGATCACCA TCAATGAGCCTAACAGGCTGAGTGACATGTACAACCGCACGAGTAATGACACCTACCGT GCAGCCCACAACCTGATGATCGCCCATGCCCAGGTCTGGCACCTCTATGATAGGCAGTA

TAGGCCGGTCCAGCATGGGGCTGTGTCGCTGTCCTTACATTGCGACTGGGCAGAACCTG CCAACCCCTTTGTGGATTCACACTGGAAGGCAGCCGAGCGCTTCCTCCAGTTTGAGATC GCCTGGTTTGCAGATCCGCTCTTCAAGACTGGCGACTATCCATCGGTTATGAAGGAATA CATCGCCTCCAAGAACCAGCGAGGGCTGTCTAGCTCAGTCCTGCCGCGCTTCACCGCGA AGGAGAGCAGGCTGGTGAAGGGTACCGTCGACTTCTACGCACTGAACCACTTCACTACG AGGTTCGTGATACACAAGCAGCTGAACACCAACCGCTCAGTTGCAGACAGGGACGTCCA

GTTCCTGCAGGACATCACCCGCCTAAGCTCGCCCAGCCGCCTGGCTGTAACACCCTGGG GAGTGCGCAAGCTCCTTGCGTGGATCCGGAGGAACTACAGAGACAGGGATATCTACATC ACAGCCAATGGCATCGATGACCTGGCTCTAGAGGATGATCAGATCCGAAAGTACTACTT GGAGAAGTATGTCCAGGAGGCTCTGAAAGCATATCTCATTGACAAGGTCAAAATCAAAG GCTACTATGCATTCAAACTGACTGAAGAGAAATCTAAGCCTAGATTTGGATTTTTCACC TCTGACTTCAGAGCTAAGTCCTCTGTCCAGTTTTACAGCAAGCTGATCAGCAGCAGTGG

CCTCCCCGCTGAGAACAGAAGTCCTGCGTGTGGTCAGCCTGCGGAAGACACAGACTGCA CCATTTGCTCATTTCTCGTGGAGAAGAAACCACTCATCTTCTTCGGTTGCTGCTTCATC TCCACTCTGGCTGTACTGCTATCCATCACCGTTTTTCATCATCAAAAGAGAAGAAAATT CCAGAAAGCAAGGAACTTACAAAATATACCATTGAAGAAAGGCCACAGCAGAGTTTTCA GCTAA (SEQ ID NO:.. 469).

The sequence of β-Klotho murine amino acids in total length (number of GenBank Accession No. NP_112457) is provided: MKTGCAAGSPGNEWIFFSSDERNTRSRKTMSNRALQRSAVLSAFVLLRAVTGFSGDGKA IWDKKQYVSPVNPSQLFLYDTFPKNFSWGVGTGAFQVEGSWKTDGRGPSIWDRYVYSHL RGVNGTDRSTDSYIFLEKDLLALDFLGVSFYQFSISWPRLFPNGTVAAVNAQGLRYYRA

LLDS LVLRNIE PIVTLYHWDL PLTLQE EYGGWKNATMIDLFNDYATYCFQTFGDRVKYW ITIHNPYLVAWHGFGTGMHAPGEKGNLTAVYTVGHNLIKAHS KVWHNYDKNFRPHQKGW LSITLGSHWIEPNRTDNMEDVINCQHSMSSVLGWFANPIHGDGDYPEFMKTGAMIPEFS EAEKEEVRGTADFFAFSFGPNNFRPSNTWKMGQNVSLNLRQVLNWIKLEYDDPQILIS ENGWFTDSYIKTEDTTAIYMMKNFLNQVLQAIKFDEIRVFGYTAWTLLDGFEWQDAYTT RRGLFYVDFNSEQKERKPKSSAHYYKQIIQDNGFPLKESTPDMKGRFPCDFSWGVTESV LKPEFTVSSPQFTDPHLYVWNVTGNRLLYRVEGVRLKTRPSQCTDYVSIKKRVEMLAKM KVTHYQFALDWTSILPTGNLSKVNRQVLRYYRCVVSEGLKLGVFPMVTLYHPTHSHLGL PLPLLSSGGWLNMNTAKAFQDYAELCFRELGDLVKLWITINEPNRLSDMYNRTSNDTYR AAHNLMIAHAQVWHLYDRQYRPVQHGAVSLSLHCDWAEPANPFVDSHWKAAERFLQFEI AWFADPLFKTGDYPSVMKEYIASKNQRGLSSSVLPRFTAKESRLVKGTVDFYALNHFTT RFVIHKQLNTNRSVADRDVQFLQDITRLSSPSRLAVTPWGVRKLLAWIRRNYRDRDIYI TANGIDDLALEDDQIRKYYLEKYVQEALKAYLIDKVKIKGYYAFKLTEEKSKPRFGFFT SDFRAKSSVQFYSKLISSSGLPAENRSPACGQPAEDTDCTICSFLVEKKPLIFFGCCFI STLAVLLSITVFHHQKRRKFQKARNLQNIPLKKGHSRVFS (SEQ ID NO:.. 468).

As described herein, β-Klotho proteins can also include fragments. As used herein, the terms are used interchangeably to mean a co-receptor, in particular, and, unless otherwise specified, a human co-receptor that, in association with FGFRlc and FGF21, induces signaling activity from the type FGF21.

The term β-Klotho also includes post-translational modifications of the β-Klotho amino acid sequence, for example, possible glycosylation sites linked to N. Thus, antigen binding proteins can bind or be generated by glycosylated proteins in one or more of the positions.

Antigen binding proteins that specifically bind to one or more of β-Klotho, FGFRlc, FGFR2c, FGFR3C and FGFR4c

Several selective binding agents useful for modulating FGF21-type signaling are provided. Such agents include, for example, antigen binding proteins that contain an antigen binding domain (for example, single chain antibodies, domain antibodies, hemibodies, immunoadhesions and polypeptides with an antigen binding region) and specifically bind to FGFRlc, β-Klotho, or both FGFRlc and β-Klotho, in particular human FGFRlc and human β-Klotho. Some of the agents, for example, are useful in mimicking the signaling effect generated in vivo by associating FGFRlc with β-Klotho and FGF21 and, thus, can be used to intensify or modulate one or more activities associated with signaling of the type FGF21.

In general, the antigen binding proteins that are provided typically comprise one or more CDRs, as described herein (for example, 1, 2, 3, 4, 5 or 6 CDRs). In some embodiments, antigen-binding proteins are naturally expressed by clones, while in other embodiments, the antigen-binding protein may comprise (a) a framework polypeptide structure and (b) one or more CDRs that are inserted and / or attached to the polypeptide structure of the framework. In some of these modalities, a CDR forms a component of heavy or light chains expressed by the clones described herein; in other modalities, a CDR can be inserted into a framework in which the CDR is not naturally expressed. A framework polypeptide structure can take many different forms. For example, a framework polypeptide structure may be, or comprise, the framework of a naturally occurring antibody, or fragment or variant thereof, or it may be of a completely synthetic nature. Examples of various antigen-binding protein structures will be further described below.

In some embodiments in which the antigen binding protein comprises (a) a framework polypeptide structure and (b) one or more CDRs that are inserted and / or joined to the framework polypeptide structure, the framework polypeptide structure of a protein antigen binding is an antibody or is derived from an antibody, including, without limitation, monoclonal antibodies, bispecific antibodies, minibodies, domain antibodies, synthetic antibodies (sometimes referred to as antibody mimetics), chimeric antibodies, humanized antibodies, fusions of antibody (sometimes called antibody conjugates), and portions or fragments of each, respectively. In some cases, the antigen binding protein is an immune fragment of an antibody (for example, a Fab, Fab ', F (ab') ₂ or scFv).

Some of the antigen binding proteins as provided herein specifically bind to: (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, including the human forms of that protein. In one embodiment, an antigen-binding protein specifically binds to both human FGFRlc which comprises the amino acid sequence of the ID. SEQ. N °: 5 for the human βKlotho that comprises the amino acid sequence of the ID. SEQ. No. 8, and in another embodiment, an antigen-binding protein specifically binds both to human FGFRlc which comprises the ID amino acid sequence. SEQ. N °: 5 for human β-Klotho that has the ID amino acid sequence. SEQ. N °: 8 and induces FGF21 type signaling. Thus, an antigen-binding protein can induce, but not necessarily, FGF21-like signaling.

Structure of the antigen binding protein

Some of the antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, including the human forms of these proteins that are provided herein, have a structure typically associated with naturally occurring antibodies. The structural units of these antibodies typically comprise one or more tetramers, each composed of two identical pairs of polypeptide chains, although some species of mammals also produce antibodies that have only a single heavy chain. In a typical antibody, each pair or doublet includes a full-length light chain (in some embodiments, about 25 kDa) and a full-length heavy chain (in certain embodiments, about 50-70 kDa). Each individual immunoglobulin chain is composed of several immunoglobulin domains, each consisting of approximately 90 to 110 amino acids and expressing a characteristic fold pattern. These domains are the basic units of which the antibody polypeptides are composed. The amino-terminal portion of each chain typically includes a variable domain that is responsible for antigen recognition. The carboxy terminal portion is more evolutionarily conserved than the other end of the chain and is called the constant region or C region. Human light chains are generally classified as kappa (K) and lambda (X) light chains, each of which contains a variable domain and a constant domain. Heavy chains are typically classified as mu, delta, gamma, alpha or epsilon chains, and these define the antibody isotype as IgM, IgD, IgG, IgA and IgE, respectively. IgG has several subtypes, including, without limitation, IgGl, IgG2, IgG3 and IgG4. IgM subtypes include IgM and IgM2. IgA subtypes include IgAl and IgA2. In humans, the IgA and IgD isotypes contain four heavy chains and four light chains; the IgG and IgE isotypes contain two heavy chains and two light chains; and the IgM isotype contains five heavy chains and five light chains. The heavy chain C region typically comprises one or more domains that may be responsible for the effector function. The number of domains in the heavy chain region will depend on the isotype. Each of the IgG heavy chains, for example, contains three domains in the C region known as C _H 1, C _H 2 and C _H 3. The antibodies that are provided can have any of these isotypes and subtypes. In certain embodiments, an antigen-binding protein that specifically binds to one or more of (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of

FGFRlc, FGFR2C, FGFR3c and FGFR4 is an antibody of the IgGl, IgG2 or IgG4 subtype.

In full-length light and heavy chains, the variable and constant regions are joined by a J region of about twelve or more amino acids, with the heavy chain also including a D region of about ten more amino acids. See, eg, Fundamental Immunology, 2nd Edition, Chapter 7 (Paul, W., ed.) , 1989, New York: Raven Press (incorporated herein by reference in its entirety for all purposes). The variable regions of each light / heavy chain pair typically form the antigen binding site.

An example of an IgG2 heavy constant domain of an exemplary monoclonal antibody that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and a FGFRlc, FGFR2c, FGFR3c and FGFR4 has the amino acid sequence: ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSWTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPS VFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TFRWSVLTWHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:.. 9).

An example of a kappa light constant domain of an exemplary monoclonal antibody that binds to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 has the amino acid sequence: RTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ

DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID. NO: 10).

An example of a light lambda constant domain of an exemplary monoclonal antibody that binds to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 has the amino acid sequence:

GQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPS KQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 11).

Variable regions of immunoglobulin chains generally exhibit the same global structure, which comprises relatively conserved framework regions (FR) joined by three hypervariable regions, more often called complementarity determining regions or CDRs. The CDRs of the two chains of each heavy chain / light chain pair mentioned above are typically aligned by the framework regions to form a structure that specifically binds to a specific epitope in the target protein (e.g., (i) β-Klotho; ( ii) FGFRlc, FGFR2c, FGFR3c or FGFR4, or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4). From terminal N to terminal C, the variable regions of the naturally occurring light and heavy chain typically conform to the following order of these elements: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. A numbering system was developed to assign numbers to the amino acids that occupy positions in each of these domains. This numbering system is defined in Kabat Sequences of Proteins of Immunological Interest (1987 and 1991, NIH, Bethesda, MD). As desired, CDRs can also be redefined according to an alternative nomenclature scheme, for example, that of Chothia (see Chothia and Lesk, 1987, J. Mol. Biol. 196: 901-917; Chothia et al., 1989 , Nature 342: 878-883 or Honegger and Pluckthun, 2001, J. Mol. Biol. 309: 657-670).

The various variable regions of the heavy chain and the light chain of antigen binding proteins provided here are revealed in Table 2. Each of these variable regions can be attached to the constant regions of the heavy and light chain above to form a heavy and light chain of complete antibody, respectively. In addition, each of the heavy and light chain sequences thus generated can be combined to form a complete antibody structure. It should be understood that the variable regions of the heavy chain and the light chain provided here can also be attached to other constant domains that have sequences different from the exemplary sequences listed above.

Specific examples of some of the full length light and heavy chains of the antibodies that are provided and their corresponding amino acid sequences are summarized in Tables IA and IB. Table IA shows exemplary light chain sequences, and Table 1B shows exemplary heavy chain sequences.

Table IA - Exemplary sequences of the antibody light chain

ID. IN SEQ. N °: Designation Contained in the clone Amino acid sequence

ID. IN SEQ. N °: Designation Contained in the clone Sequence of amino acids 12 LI 17C3 SYVLTQPPSVSVAPGQTARITCGGNNIGSQSV HWYQQKPGQAPVLWYDDSDRPSGI PERFSGS NSGNTATLTISRVEAGDEADYYCQVWDSSSDH WFGGGTKLTVLGQPKANPTVTLFPPSSEELQ ANKATLVCLISDFYPGAVTVAWKADGSPVKAG VETTKPSKQSNNKYAASSYLSLTPEQWKSHRS YSCQVTHEGSTVEKTVAPTECS 13 L2 22H5 SYVLTQPPSVSVAPGQTARITCGGNNIGSQSV HWYQQKPGQAPVLWYDDSDRPSGI PERFSGS NSGNTATLTISRVEAGDEADYYCQVWDNTSDH WFGGGTKLTVLGQPKANPTVTLFPPSSEELQ ANKATLVCLISDFYPGAVTVAWKADGS PVKAG VETTKPSKQSNNKYAASSYLSLTPEQWKSHRS YSCQVTHEGSTVEKTVAPTECS 14 L3 16H7 24H11 SYVLTQPPSVSVAPGQTARITCGGNNIGSESV HWYQQKPGQAPVLWYDDSDRPSGI PERFSGS NSGNTATLTISRVEAGDEADYYCQVWDGNSDH WFGGGTKLTVLGQPKANPTVTLFPPSSEELQ ANKATLVCLISDFYPGAVTVAWKADGS PVKAG VETTKPSKQSNNKYAASSYLSLTPEQWKSHRS YSCQVTHEGSTVEKTVAPTECS 15 L4 18G1 EIVLTQSPGTLSLSPGERATLSCRASQNFDSS YLAWYQQKPGQAPRLLIYGTSSRATGIPDRFS GIGSGTDFTLTINRLEPEDFAMYYCQQYGGSP LTFGGGTEVEIKRTVAAPSVFIFPPSDEQLKS GTASWCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKV

ID. IN SEQ. N °: Designation Contained in the clone Amino acid sequence YACEVTHQGLS S PVTKS FNRGEC 16 L5 17D8 EIVLTQSPGTLSLSPGERATLSCRASQSVSGN YLAWYQQKPGQAPRLLIYGASSRATGIPDRFS GSGSGTDFTLTISRLEPEDFAVYYCQQYGSAP LTFGGGTKVEIKRTVAAPSVFIFWPSKLQKTKKLQKTKKLQKKLQQKKQQ YACEVTHQGLS S PVTKS FNRGEC 17 L6 26H11 EIVLTQSPGTLSLSPGERATLSCRASQSVSGN YLAWYQQKPGQAPRLLIYGASSRATGIPDRFS GSGSGTDFTLTISRLEPEDFAMYYCQQYGSSP LTFGGGSKVEIKRTVAAPSVFIFWPSKLQSKESQSQSKLQSKLQKKLQKKQQ YACEVTHQGLS S PVTKS FNRGEC 18 L7 12E4 EIVLTQSPGTLSLSPGERATLSCRASQNFDSN YLAWYQQKPGQAPRLLIYGASSRATGIPDNFS GSGSGTDFTLTISRLEPEDFAMYYCQQYGSSP LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKS GTASWCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLS S PVTKS FNRGEC 19 L8 12C11 EIVLTQSPGTLSLSPGERATLSCRASQNFDSS SLAWYQQKPGQAPRLLIYGASSRATGIPDRFS GSGSGTDFTLTISRLEPEDFAMYYCQQCGSSP LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKS GTASWCLLNNFYPREAKVQWKVDNALQSGNS

ID. IN SEQ. N °: Designation Contained in the clone Amino acid sequence QESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGL S S PVTKS FNRGEC 20 L9 21H2 21B4 EIVLTQS PGTLSLS PGERATLS CRASQSVS ST YLAWHQQKPGQGLRLLIYGASSRATGIPDRFS GSGSGTDFTLTISRLEPEDFAVYYCQQYGSSF TFGGGTRVEIKRTVAAPSVFIFPPSDEQLKSG TASWCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGL S PVTKS FNRGEC 21 IOL 18B11.1 DIVMTQSPLSLPVTPGEPASISCRSSQSLLYY NGFTYLDWFLQKPGQS PHLLIYLGSNRASGVP DRFSGSVSGTDFTLKISRVEAEDVGVYYCMQS LQTPFTFGPGTKVDIKRTVAAPSVFIFPPSDE QLKSGTASWCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLS S PVTKS FNRGEC 22 Lil 18B11.2 EIVMTQSPATLSVSPGERATLSCRASQSVNSN LAWYQQKPGQAPRLLIYGVSTRATGIPARFSG SGSGTEFTLTIRSLQSEDFAVYYCQQYNNWPP TFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSG TASWCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLS S PVTKS FNRGEC 23 L12 2 0D4 DIQLTQSPSSLSASIGDRVTITCRASQDIRYD LGWYQQKPGKAPKRLIYAASSLQSGVPSRFSG SGSGTEFTLTVSSLQPEDFATYYCLQHNSYPL TFGGGTKVEIERTVAAPSVFIFPPSDEQLKSG

ID. IN SEQ. N °: Designation Contained in the clone Amino acid sequence TASWCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLS S PVTKS FNRGEC 24 L13 46D11 DIQMTQSPSSVSASVGDRVTITCRASQGISIW LAWYQQKPGKAPKLLIYAASSLQSGVPSRFSG SGSGTDFTLTISSLQPEDFATYYCQQANDFPI TFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSG TASWCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLS S PVTKS FNRGEC 25 L14 4 0D2 DFVMTQTPLSLSVTPGQPASISCKSSQSLLQS DGKTYLYWYLQKPGQPPHLLIYEVSNRFSGVP DRFSGSGSGTDFTLKISRVEAEDVGVYYCMQS IQLPRTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASWCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGL S PVTKS FNRGE C 26 L15 37D3 DIVMTQSPLSLPVTPGEPASISCRSSQSLLHS NGYNFLDWYLQKPGQSPQLLIYLGSDRASGVP DRFSGSGSGTEFTLKISRVEAEDVGLYYCMQA LQTPCSFGQGTKLEIKRTVAAPSVFIFPPSDE QLKSGTASWCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGEC 27 L16 39F7 EIVLTQSPGTLSLSPGERATLSCRASQSVSST YLAWYQQKPGQAPRLLIYGAS SRATGIPDRFS GSGSGTDFTLTISRLEPEDFAVYYCQQSGSSP

ID. IN SEQ. N °: Designation Contained in the clone Amino acid sequence LTFGGGTEVEIKRTVAAPSVFIFPPSDEQLKS GTASWCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLS S PVTKS FNRGEC 28 L17 39F11 EIVLTQSPGTLSLSPGERATLSCRASQSVSST YLAWYQQKPGQAP S LLIYGAS SRATGIPDRFS GSGSGTDFTLTISRLEPEDFAVYYCQQSGSSP LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKS GTASWCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLS S PVTKS FNRGEC 29 L18 3 9G5 EIVLTQSPGTLSLSPGERATLSCRASQSVSST YLAWYQQKPGQAPRLLIYGASFRATGIPDRFS GSGSGTDFTLTISRLEPEDFAVYYCQQSGSSP LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKS GTASWCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLS S PVTKS FNRGE C

Table IB - Exemplary sequences of the heavy antibody chain

ID. IN SEQ. N °: Designation Contained in the clone Sequence 30 H1 17C3 QVTLKESGPVLVKPTETLTLTCTVSGFSLSNA RMGVSWIRQPPGKALEWLAHIFSNDEKSYSTS LKS RLTIS KDTS KSQWLTMTNMDP VDTAT Y Y CARILLLGAYYYYGMDVWGQGTTVSTSSFSGSVTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTLFLKLFLFLFLFLFLLFLLLLLLLLLL

ID. IN SEQ. N °: Designation Contained in the clone Sequence VTVPSSNFGTQTYTCNVDHKPSNTKVDKTVER KCCVECPPCPAPPVAGPSVFLFPPKPKDTLMI SRTPEVTCVWDVSHEDPEVQFNWYVDGVEVH NAKTKPREEQFNSTFRWSVLTVVHQDWLNGK EYKCKVSNKGLPAPIEKTISKTKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPMLDSDGSFFLYSKLTVD KS RWQQGNVFS P LS LS GK CSVMHEALHNHYTQKS 31 H2 22H5 QVTLKESGPVLVKPTETLTLTCTVSGFSLSNA RMGVSWIRQPPGKALEWLAHIFSNDEKSYSTS LKS RLTIS KDTS WLTMTNMD KS Q P Y Y VDTAT CARILLVGAYYYCGMDVWGQGTTVTVSSASTK GPSVFPLAPCSRSTSESTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSNFGTQTYTCNVDHKPSNTKVDKTVER KCCVECPPCPAPPVAGPSVFLFPPKPKDTLMI SRTPEVTCVWDVSHEDPEVQFNWYVDGVEVH NAKTKPREEQFNSTFRVVSVLTWHQDWLNGK EYKCKVSNKGLPAPIEKTISKTKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPMLDSDGSFFLYSKLTVD GK KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP

ID. IN SEQ. N °: Designation Contained in the clone Sequence 32 H3 16H7 QVTLKESGPVLVKPTETLTLTCTVSGFSLNNA RMGVSWIRQPPGKALEWLAHIFSNDEKSYSTS LKSRLTISKDTSKSQWLIMTNMDPVDTATYY CARSWTGGYYYDGMDVWGQGTTVTVSSASTK GPSVFPLAPCSRSTSESTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSNFGTQTYTCNVDHKPSNTKVDKTVER KCCVECPPCPAPPVAGPSVFLFPPKPKDTLMI SRTPEVTCWVDVSHEDPEVQFNWYVDGVEVH NAKTKPREEQFNSTFRWSVLTWHQDWLNGK EYKCKVSNKGLPAPIEKTISKTKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPMLDSDGSFFLYSKLTVD KS RWQQGNVFS CSVMHEALHNHYTQKS L S P LS GK 33 H4 24H11 QVTLKESGPVLVKPTETLTLTCTVSGFSLSNA RMGVSWIRQPPGKALEWLAHIFSNDEKSYSTS LKNRLTIS KDTS WLIMTNMD KS Q P Y Y VDTAT CARSWTGGYYYDGMDVWGQGTTVTVSSASTK GPSVFPLAPCSRSTSESTAALGCL.VKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSNFGTQTYTCNVDHKPSNTKVDKTVER KCCVECPPCPAPPVAGPSVFLFPPKPKDTLMI SRTPEVTCVWDVSHEDPEVQFNWYVDGVEVH NAKTKPREEQFNSTFRWSVLTWHQDWLNGK EYKCKVSNKGLPAPIEKTISKTKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPMLDSDGSFFLYSKLTVD GK KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP 34 H5 18G1 EVQLLESGGGLVQPGGSLRLSCAASRFTFSTY AMSWVRQAPGKGLEWVSGISGSGVSTHYADSV KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKSLIWIVYALDHWGQGTLVTVSSASTKGPS VFPLAPCSRSTSESTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSWTV PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCC VECPPCPAPPVAGPSVFLFPPKPKDTLMISRT

ID. IN SEQ. N °: Designation Contained in the clone Sequence PEVTCWVDVSHEDPEVQFNWYVDGVEVHNAK TKPREEQFNSTFRWSVLTWHQDWLNGKEYK CKVSNKGLPAPIEKTISKTKGQPREPQVYTLP PSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPMLDSDGSFFLYSKLTVDKSR WQQGNVFS CSVMHEALHNHYTQKS L S L S PGK 35 H6 17D8 EVQLLESGGGLVQPGGYLRLSCAASGFTFSTY AMSWVRQAPGKGLEWVSAISGSGVSTYYADSV KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKSLIWMVYVLDYWGQGTLVTVSSASTKGPS VFPLAPCSRSTSESTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSWTV PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCC VECPPCPAPPVAGPSVFLFPPKPKDTLMISRT PEVTCWVDVSHEDPEVQFNWYVDGVEVHNAK TKPREEQFNSTFRWSVLTWHQDWLNGKEYK CKVSNKGLPAPIEKTISKTKGQPREPQVYTLP PSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPMLDSDGSFFLYSKLTVDKSR WQQGNVFS CSVMHEALHNHYTQKS LS U S PGK 36 H7 26H11 EVQLLESGGGLVQPGGYLRLSCAASGFTFSTY AMSWVRQAPGKGLEWVSAISGSGVSTNYADSV KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKS LIWMVYVLDYWGQGTLVTVS SAS TKGPS VFPLAPCSRSTSESTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSWTV PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCC VECPPCPAPPVAGPSVFLFPPKPKDTLMISRT PEVTCWVDVSHEDPEVQFNWYVDGVEVHNAK TKPREEQFNSTFRWSVLTWHQDWLNGKEYK CKVSNKGLPAPIEKTISKTKGQPREPQVYTLP PSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPMLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 37 H8 12E4 12C11 EVQLLESGGGLVQPGGSLRLSCAASRFTFSTY AMSWVRQAPGKGLEWVSGISGSGVSTYYADSV KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKSLIWIVYALDYWGQGTLVTVSSASTKGPS

ID. IN SEQ. N °: Designation Contained in the clone Sequence VFPLAPCSRSTSESTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSWTV PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCC VECPPCPAPPVAGPSVFLFPPKPKDTLMISRT PEVTCWVDVSHEDPEVQFNWYVDGVEVHNAK TKPRE T EQ FNS FRWS VLT WHQDWLNGKE YK CKVSNKGLPAPIEKTIS KTKGQPRE PQVYTLP PSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPMLDSDGSFFLYSKLTVDKSR WQQGNVF CSVMHEALHNHYTQKS S S L LS PGK 38 H9 21H2 QVQLQESGPGLVKPSETLSLTCTVSGGSISSY YWSWIRQPAGKGLEWIGRIYTSGSTNYNPSLK SRVTMSKDTSKNQFSLKLRSVTAADTAVYYCA RDPDGDYYYYGMDVWGQGTSVTVSSASTKGPS VFPLAPCSRSTSESTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSWTV PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCC VECPPCPAPPVAGPSVFLFPPKPKDTLMISRT PEVTCWVDVSHEDPEVQFNWYVDGVEVHNAK TKPRE EQ FNS T FRWS VLT WHQDWLNGKE YK CKVSNKGLPAPIEKTISKTKGQPREPQVYTLP PSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPMLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 39 H10 21B4 QVQLQESGPGLVKPSETLSLTCTVSGGSISSY FWSWIRQPAGKGLEWIGRIYTSGSTNYNPSLK SRVTMSIDTSKNQFSLKLSSVTAADTAVYYCA RDPDGDYYYYGMDVWGQGTTVTVSSASTKGPS VFPLAPCSRSTSESTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSWTV PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCC VECPPCPAPPVAGPSVFLFPPKPKDTLMISRT PEVTCWVDVSHEDPEVQFNWYVDGVEVHNAK TKPREEQFNSTFRWSVLTWHQDWLNGKEYK CKVSNKGLPAPIEKTISKTKGQPREPQVYTLP PSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPMLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

ID. IN SEQ. N °: Designation Contained in the clone Sequence 40 Hll 18B11.1 18B11.2 EVQLVESGGGLVKPGGSLRLSCAASGFTFSDA WMSWVRQAPGKGLEWVGRIKSKTDGGTTDYAA PVKGRFTISRDDSKNTLYLQMNSLKTEDTAVY F CTS TYS S GWYVWDYYGMDVWGQGTTVTVS SA STKGPSVFPLAPCSRSTSESTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSWTVPSSNFGTQTYTCNVDHKPSNTKVDKT VERKCCVECPPCPAPPVAGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVQFNWYVDGV EVHNAKTKPREEQFNSTFRVVSVLTVVHQDWL NGKEYKCKVSNKGLPAPIEKTISKTKGQPREP QVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPMLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPGK 41 H12 20D4 QVQLVQSGAEVKKPGASVKVSCKVSGYTLTDL SMHWVRQAPGKGLEWMGGFDPEDGETIYAQKF QGRITMTEDTSTDTAYMELSSLRSEDTAVYYC ASIWVPAAIQSYYYYYGMGVWGQGTTVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDY FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS LSSWTVPSSNFGTQTYTCNVDHKPSNTKVDK TVERKCCVECPPCPAPPVAGPSVFLFPPKPKD TLMISRTPEVTCVWDVSHEDPEVQFNWYVDG VEVHNAKTKPREEQFNSTFRWSVLTWHQDW LNGKEYKCKVSNKGLPAPIEKTISKTKGQPRE PQVYTLPPSREEMTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPMLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK 42 H13 46D11 QVTLKEAGPVLVKPTETLTLTCTVSGFSLSNA RMGVNWIRQPPGKALEWLAHIFSNDEKSYSTS LKSRLTISKDTSKSQWLTMTNMDPVDTATYY CARVRIAGDYYYYYGMDVWGQGTTVTVSSAST KGPSVFPLAPCSRSTSESTAALGCLVKDYFPE PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS WTVPSSNFGTQTYTCNVDHKPSNTKVDKTVE RKCCVECPPCPAPPVAGPSVFLFPPKPKDTLM

ID. IN SEQ. N °: Designation Contained in the clone Sequence ISRTPEVTCVWDVSHEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTFRWSVLTWHQDWLNG KEYKCKVSNKGLPAPIEKTISKTKGQPREPQV YTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPMLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK 43 H14 39F11 QVQLVESGGGWQPGRSLRLSCAASGFTFSSY GIHWVRQAPGKGLEWVAVIWYDGSDKYYADSV KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC ARDRAAAGLHYYYGMDVWGQGTTVTVSSASTK GPSVFPLAPCSRSTSESTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSNFGTQTYTCNVDHKPSNTKVDKTVER KCCVECPPCPAPPVAGPSVFLFPPKPKDTLMI SRTPEVTCWVDVSHEDPEVQFNWYVDGVEVH NAKTKPREEQ FNS T FRWS VLT WHQDWLNGK EYKCKVSNKGLPAPIEKTIS KTKGQPRE PQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPMLDSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK 44 H15 39F7 QVQLVESGGGWQPGRSLRLSCAASGFTFSNY GIHWVRQAPGKGLEWVAVIWYDGSIKYYADSV KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC ARDRAAAGLHYYYGMDVWGQGTTVTVS are the TK GPSVFPLAPCSRSTSESTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSNFGTQTYTCNVDHKPSNTKVDKTVER KCCVECPPCPAPPVAGPSVFLFPPKPKDTLMI SRTPEVTCVWDVSHEDPEVQFNWYVDGVEVH NAKTKPREEQFNSTFRWSVLTWHQDWLNGK EYKCKVSNKGLPAPIEKTISKTKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPMLDSDGSFFLYSKLTVD GK KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP

ID. IN SEQ. N °: Designation Contained in the clone Sequence 45 H16 39G5 QVQLVESGGGWQPGRSLRLSCAVSGFTFSSY GIHWVRQAPGKGLEWVAVIWYDGSDKYYGDSV KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC ARDRAAAGLHYYYGMDVWGQGTTVTVSSASTK GPSVFPLAPCSRSTSESTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSNFGTQTYTCNVDHKPSNTKVDKTVER KCCVECPPCPAPPVAGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVH NAKTKPREEQFNSTFRWSVLTWHQDWLNGK EYKCKVSNKGL PAPIEKTISKTKGQPRE PQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPMLDSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK 46 H17 40D2 QVQLQESGPGLVKPSQTLSLTCTVSGGSISSG GYNWSWIRQHPGKGLEWIGNIYYSGSTYYNPS LKSRVTISVDTSKNQFSLKLRSVTAADTAVYY CARENIWIPAAIFAGWFDPWGQGTLVTVS SA STKGPSVFPLAPCSRSTSESTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSWTVPSSNFGTQTYTCNVDHKPSNTKVDKT VERKCCVECPPCPAPPVAGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVQFNWYVDGV EVHNAKTKPREEQFNSTFRWSVLTWHQDWL NGKEYKCKVSNKGLPAPIEKTISKTKGQPREP QVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPMLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPGK 47 H18 37D3 EVHLVESGGGLAKPGGSLRLSCAASGFTFRNA WMSWVRQAPGKGLEWVGRIKSKTDGGTTDYAA PVKGRFTISRDDSKNTLYLQMNSLKTEDTAEY YCITDRVLSYYAMAVWGQGTTVTVS SASTKGP SVFPLAPCSRSTSESTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSWT VPSSNFGTQTYTCNVDHKPSNTKVDKTVERKC CVECPPCPAPPVAGPSVFLFPPKPKDTLMISR

ID. IN SEQ. N °: Designation Contained in the clone Sequence TPEVTCVWDVSHEDPEVQFNWYVDGVEVHNA KTKPRE EQ FNS TFRWS VLT WHQDWLNGKE Y KCKVSNKGLPAPIEKTISKTKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPMLDSDGSFFLYSKLTVDKS RWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

Again, each of the exemplary heavy chains (H1, H2, H3 etc.) listed in Tables IB and 6A, infra, can be combined with any of the exemplary light chains shown in Tables IA and 6A, infra, to form an antibody. Examples of such combinations include H1 combined with any one of LI to L18; H2 combined with any one of LI to L18; H3 combined with any one of LI to L18, and so on. In some cases, antibodies include at least one heavy and one light chain from those listed in Tables IA and IB and 6A, infra; particular examples of light and heavy chain pairings include LI with HI, L2 with H2, L3 with H3, L4 with H4, L5 with H5, L6 with H6, L7 with H7, L8 with H8, L9 with H9, LIO with H10 , Lll with Hll, L12 with H12, L13 with H13, L14 with H14, L15 with H15, L16 with H16, L17 with H17 and L18 with H18. In addition to antigen-binding proteins that comprise a heavy chain and a light chain from the same clone, a heavy chain from a first clone can be paired with a light chain from a second clone (for example, a 46D11 heavy chain paired with a 16H7 light chain or a 16H7 heavy chain paired with a 46D11 light chain). These pairings can generally include VL with

90% or more homology that can be paired with the naturally occurring clone heavy chain. In some cases, the antibodies comprise two different heavy chains and two different light chains listed in Tables IA and 1B and 6A, infra. In other cases, the antibodies contain two identical light chains and two identical heavy chains. As an example, an immunologically functional antibody or fragment may include two H1 heavy chains and two L1 light chains, or two H2 heavy chains and two L2 light chains, or two H3 heavy chains and two L3 light chains, and other similar combinations of pairs of light chains and heavy chain pairs, as listed in Tables IA and IB and 6A, infra.

In another aspect of the present disclosure, hemibodies are provided. A hemibody is a monovalent antigen binding protein comprising (i) an intact light chain, and (ii) a heavy chain fused to an Fc region (for example, an IgG2 Fc region of SEQ ID. 441), optionally via a linker. The linker can be a linker (G ₄ S) _X , where x is an integer other than zero (for example, (G ₄ S) ₈ ; SEQ ID. No.: 440). Hemibodies can be built using the supplied heavy and light chain components. Specific examples of hemibodies are disclosed in Example 14.

Other antigen binding proteins that are provided are antibody variants formed by combining the heavy and light chains shown in Tables IA and IB and 6A, infra, and comprise light and / or heavy chains that each have at least 70 %, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity for the amino acid sequences of these chains. In some cases, these antibodies include at least one heavy chain and one light chain, while in other cases, the variant forms contain two identical light chains and two identical heavy chains.

Variable domains of antigen binding proteins

Antigen binding proteins are also provided that contain a variable region of the antibody heavy chain selected from the group consisting of V _H 1, V _H 2, V _h 3, V _h 4, V _h S, V _h 6, V _h 7, V _h 8, V _H 9, V _H 10, V _H 11, V _H 12, V _H 13, V _h 14, V _h 15, V _h 16, V _h 17 and V _H 18, as shown in Table 2B, and / or a variable region of the antibody light chain selected from the group consisting of V _L 1, V _L 2, V _L 3, V _L 4, V _L S, V _l 6, V _l 7, V _l 8 , V _l 9, V _l 10, V _l 11, V _l 12, V _L 13, V _l 14, V _L 15, V _l 16, V _l 17 and V _l 18, as shown in Table 2A, and fragments immunologically functional, derivatives, muteins and variants of these variable regions of light chain and heavy chain.

Table 2A - Exemplary variable light chains (V _L ) of antibody.

Contained in the clone Designation ID. IN SEQ. N °: Sequence of amino acids 17C3 V _L 1 48 SYVLTQPPSVSVAPGQTARITCGGNNIGSQ SVHWYQQKPGQAPVLWYDDSDRPSGI PER FSGSNSGNTATLTISRVEAGDEADYYCQVW DSSSDHWFGGGTKLTVL 22H5 V _L 2 49 SYVLTQPPSVSVAPGQTARITCGGNNIGSQ SVHWYQQKPGQAPVLWYDDSDRPSGI PER FSGSNSGNTATLTISRVEAGDEADYYCQVW DNTSDHWFGGGTKLTVL

Contained in the clone Designation ID. IN SEQ. N °: Sequence of amino acids 16H7 24H11 V _l 3 50 SYVLTQPPSVSVAPGQTARITCGGNNIGSE SVHWYQQKPGQAPVLWYDDSDRPSGI PER FSGSNSGNTATLTISRVEAGDEADYYCQVW DGNSDHWFGGGTKLTVL 18G1 V _l 4 51 EIVLTQS PGTLSLS PGERATLS CRASQNFD S SYLAWYQQKPGQAPRLLIYGTS SRATGIP DRFSGIGSGTDFTLTINRLEPEDFAMYYCQ QYGGS PLTFGGGTEVEIK 17D8 V _l 5 52 EIVLTQSPGTLSLSPGERATLSCRASQSVS GNYLAWYQQKPGQAPRLLIYGASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVYYCQ QYGSAPLTFGGGTKVEIK 26H11 V _l 6 53 EIVLTQSPGTLSLSPGERATLSCRASQSVS GNYLAWYQQKPGQAPRLLIYGASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFAMYYCQ QYGSSPLTFGGGSKVEIK 12E4 V _l 7 54 EIVLTQSPGTLSLSPGERATLSCRASQNFD SNYLAWYQQKPGQAPRLLIYGASSRATGIP DNFSGSGSGTDFTLTISRLEPEDFAMYYCQ QYGS S PLTFGGGTKVEIK 12C11 V _l 8 55 EIVLTQSPGTLSLSPGERATLSCRASQNFD S S S LAWYQQKPGQAPRLLIYGASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFAMYYCQ QCGSSPLTFGGGTKVEIK 21H2 21B4 V _l 9 56 EIVLTQSPGTLSLSPGERATLSCRASQSVS S TYLAWHQQKPGQGLRLLIYGASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVYYCQ QYGS S FTFGGGTRVEIK

Contained in the clone Designation ID. IN SEQ. N °: Sequence of amino acids 18B11.1 V _L 10 57 DIVMTQSPLSLPVTPGEPASISCRSSQSLL YYNGFTYLDWFLQKPGQS PHLLIYLGSNRA SGVPDRFSGSVSGTDFTLKISRVEAEDVGV YYCMQSLQTPFTFGPGTKVDIK 18B11.2 V _L 11 58 EIVMTQSPATLSVSPGERATLSCRASQSVN SNLAWYQQKPGQAPRLLIYGVSTRATGIPA RFSGSGSGTEFTLTIRSLQSEDFAVYYCQQ YNNWPPTFGQGTKVEIK 2 0D4 V _l 12 59 DIQLTQSPSSLSASIGDRVTITCRASQDIR YDLGWYQQKPGKAPKRLIYAASSLQSGVPS RFSGSGSGTEFTLTVSSLQPEDFATYYCLQ HNSYPLTFGGGTKVEIE 46D11 V _l 13 60 DIQMTQSPSSVSASVGDRVTITCRASQGIS IWLAWYQQKPGKAPKLLIYAASSLQSGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCQQ ANDF PITFGQGTRLEIK 40D2 V _l 14 61 DFVMTQTPLSLSVTPGQPASISCKSSQSLL QSDGKTYLYWYLQKPGQPPHLLIYEVSNRF SGVPDRFSGSGSGTDFTLKISRVEAEDVGV YYCMQSIQLPRTFGQGTKVEIK 37D3 V _l 15 62 DIVMTQSPLSLPVTPGEPASISCRSSQSLL HSNGYNFLDWYLQKPGQS PQLLIYLGSDRA SGVPDRFSGSGSGTEFTLKISRVEAEDVGL YYCMQALQTPCSFGQGTKLEIK 39F7 V _l 16 63 EIVLTQSPGTLSLSPGERATLSCRASQSVS STYLAWYQQKPGQAPRLLIYGASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVYYCQ QSGS S PLTFGGGTEVEIK

Contained in the clone Designation ID. IN SEQ. N °: Sequence of amino acids 39F11 V _l 17 64 EIVLTQSPGTLSLSPGERATLSCRASQSVS STYLAWYQQKPGQAPSLLIYGASSRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVYYCQ QSGSS PLTFGGGTKVEIK 39G5 V _l 18 65 EIVLTQSPGTLSLSPGERATLSCRASQSVS S TYLAWYQQKPGQAPRLLIYGAS FRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVYYCQ QSGSSPLTFGGGTKVEIK

Table 2B - Exemplary variable heavy chains (V _H ) of antibody.

Contained in the clone Designation ID. IN SEQ. N °: Sequence of amino acids 17C3 V _H 1 66 QVTLKESGPVLVKPTETLTLTCTVSGFSLS NARMGVSWIRQPPGKALEWLAHIFSNDEKS YSTSLKSRLTISKDTSKSQWLTMTNMDPV DTATYYCARILLLGAYYYYGMDVWGQGTTV TVSS 22H5 V _h 2 67 QVTLKESGPVLVKPTETLTLTCTVSGFSLS NARMGVSWIRQPPGKALEWLAHIFSNDEKS YSTSLKSRLTISKDTSKSQWLTMTNMDPV DTATYYCARILLVGAYYYCGMDVWGQGTTV TVSS 16H7 V _h 3 68 QVTLKESGPVLVKPTETLTLTCTVSGFSLN NARMGVSWIRQPPGKALEWLAHIFSNDEKS YSTSLKSRLTISKDTSKSQWLIMTNMDPV DTATYYCARSWTGGYYYDGMDVWGQGTTV TVSS 24H11 V _h 4 69 QVTLKESGPVLVKPTETLTLTCTVSGFSLS

Contained in the clone Designation ID. IN SEQ. N °: Sequence of amino acids NARMGVSWIRQPPGKALEWLAHIFSNDEKS YSTSLKNRLTISKDTSKSQWLIMTNMDPV DTATYYCARSWTGGYYYDGMDVWGQGTTV TVSS 18G1 V _h 5 70 EVQLLESGGGLVQPGGSLRLSCAASRFTFS TYAMSWVRQAPGKGLEWVSGISGSGVSTHY ADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKSLIWIVYALDHWGQGTLVTVS s 17D8 V _h 6 71 EVQLLESGGGLVQPGGYLRLSCAASGFTFS TYAMSWVRQAPGKGLEWVSAISGSGVSTYY ADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKSLIVVMVYVLDYWGQGTLVTVS S 26H11 V _h 7 72 EVQLLESGGGLVQPGGYLRLSCAASGFTFS TYAMSWVRQAPGKGLEWVSAISGSGVSTNY ADSVKGRFTISRDNS KNTLYLQMNSLRAED TAVYYCAKS LIVVMVYVLDYWGQGTLVTVS S 12E4 12C11 V _h 8 73 EVQLLESGGGLVQPGGSLRLSCAASRFTFS TYAMSWVRQAPGKGLEWVS GISGSGVSTYY ADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKS LIWIVYALDYWGQGTLVTVS S 21H2 V _h 9 74 QVQLQESGPGLVKPSETLSLTCTVSGGSIS SYYWSWIRQPAGKGLEWIGRIYTSGSTNYN PSLKSRVTMSKDTSKNQFSLKLRSVTAADT AVYYCARDPDGDYYYYGMDVWGQGTSVTVS

Contained in the clone Designation ID. IN SEQ. N °: Sequence of amino acids s 21B4 V _H 10 75 QVQLQESGPGLVKPSETLSLTCTVSGGSIS SYFWSWIRQPAGKGLEWIGRIYTSGSTNYN PSLKSRVTMSIDTSKNQFSLKLSSVTAADT AVYYCARDPDGDYYYYGMDVWGQGTTVTVS s 18B11.1 V _h 11 76 EVQLVESGGGLVKPGGSLRLSCAASGFTFS DAWMSWVRQAPGKGLEWVGRIKSKTDGGTT DYAAPVKGRFTISRDDSKNTLYLQMNSLKT EDTAVYF CTS TYS S GWYVWDYYGMDVWGQG TTVTV 18B11.2 V _h 11 77 EVQLVESGGGLVKPGGSLRLSCAASGFTFS DAWMSWVRQAPGKGLEWVGRIKSKTDGGTT DYAAPVKGRFTISRDDSKNTLYLQMNSLKT EDTAVYF CTS TYS S GWYVWDYYGMDVWGQG TTVTV 2 0D4 V _h 12 78 QVQLVQSGAEVKKPGASVKVSCKVSGYTLT DLSMHWVRQAPGKGLEWMGGFDPEDGETIY AQKFQGRITMTEDTSTDTAYMELSSLRSED TAVYYCASIVWPAAIQS YYYYYGMGVWGQ GTTVTVSS 46D11 V _h 13 79 QVTLKEAGPVLVKPTETLTLTCTVSGFSLS NARMGVNWIRQPPGKALEWLAHIFSNDEKS YSTSLKSRLTISKDTSKSQWLTMTNMDPV DTATYYCARVRIAGDYYYYYGMDVWGQGTT VTVSS 40D2 V _h 14 80 QVQLQESGPGLVKPSQTLSLTCTVSGGSIS SGGYNWSWIRQHPGKGLEWIGNIYYSGSTY

Contained in the clone Designation ID. IN SEQ. N °: Sequence of amino acids YNPSLKSRVTISVDTSKNQFSLKLRSVTAA DTAVYYCARENIWIPAAIFAGWFDPWGQG TLVTVSS 37D3 V _h 15 81 EVHLVESGGGLAKPGGSLRLSCAASGFTFR NAWMSWVRQAPGKGLEWVGRIKSKTDGGTT DYAAPVKGRFTISRDDSKNTLYLQMNSLKT EDTAEYYCITDRVLSYYAMAVWGQGTTVTV SS 39F7 V _h 16 82 QVQLVESGGGWQPGRSLRLSCAASGFTFS NYGIHWVRQAPGKGLEWVAVIWYDGSIKYY ADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCARDRAAAGLHYYYGMDVWGQGTTV TVSS 39F11 V _h 17 83 QVQLVESGGGWQPGRSLRLSCAASGFTFS SYGIHWVRQAPGKGLEWVAVIWYDGSDKYY ADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCARDRAAAGLHYYYGMDVWGQGTTV TVSS 39G5 V _h 18 84 QVQLVESGGGWQPGRSLRLSCAVSGFTFS SYGIHWVRQAPGKGLEWVAVIWYDGSDKYY GDSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCARDRAAAGLHYYYGMDVWGQGTTV TVSS

Table 2C - Coding sequence for variable light chains (V _L ) of antibody.

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence 17C3 V _L 1 85 TCCTATGTGCTGACTCAGCCACCCTCGGTG

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence TCAGTGGCCCCAGGTCAGACGGCCAGGATT ACCTGTGGGGGAAACAACATTGGAAGTCAG AGTGTGCACTGGTACCAGCAGAAGCCAGGC CAGGCCCCTGTCCTGGTCGTCTATGATGAT AGCGACCGGCCCTCAGGGATCCCTGAGCGA TTCTCTGGCTCCAACTCTGGGAACACGGCC ACCCTGACCATCAGCAGGGTCGAAGCCGGG GATGAGGCCGACTATTACTGTCAGGTGTGG GATAGTAGTAGTGATCATGTGGTATTCGGC GGAGGGACCAAGCTGACCGTCCTA 22H5 V _l 2 86 TCCTATGTGCTGACTCAGCCACCCTCGGTG TCAGTGGCCCCAGGACAGACGGCCAGGATT ACCTGTGGGGGAAACAACATTGGAAGTCAA AGTGTGCACTGGTACCAGCAGAAGCCAGGC CAGGCCCCTGTCCTGGTCGTCTATGATGAT AGCGACCGGCCCTCAGGGATCCCTGAGCGA TTCTCTGGCTCCAACTCTGGGAACACGGCC ACCCTGACCATCAGCAGGGTCGAAGCCGGG GATGAGGCCGACTATTACTGTCAGGTGTGG GATAATACTAGTGATCATGTGGTATTCGGC GGGGGGACCAAACTGACCGTCCTA 16H7 24H11 V _l 3 87 TCCTATGTGCTGACTCAGCCACCCTCGGTG TCAGTGGCCCCAGGACAGACGGCCAGGATT ACCTGTGGGGGAAACAACATTGGAAGTGAA AGTGTGCACTGGTACCAGCAGAAGCCAGGC CAGGCCCCTGTGCTGGTCGTCTATGATGAT AGCGACCGGCCCCTAGGGGGA

100

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence ACCCTGACCATCAGCAGGGTCGAAGCCGGG GATGAGGCCGACTATTACTGTCAGGTGTGG GATGGTAATAGTGATCATGTGGTATTCGGC GGAGGGACCAAGCTGACCGTCCTA 18G1 V _l 4 88 GAAATTGTGTTGACGCAGTCTCCAGGCACC CTGTCTTTGTCTCCAGGGGAAAGAGCCACC CTCTCCTGCAGGGCCAGTCAGAATTTTGAC AGCAGTTACTTAGCCTGGTACCAGCAGAAA CCTGGCCAGGCTCCCCGGCTCCTCATCTAT GGTACATCCAGCAGGGCCACTGGCATCCCA GACAGGTTCAGTGGCATTGGGTCTGGGACA GACTTCACTCTCACCATCAACAGACTGGAG CCTGAAGATTTTGCAATGTATTACTGTCAG CAGTATGGTGGCTCACCGCTCACTTTCGGC GGAGGGACCGAGGTGGAAATCAAA 17D8 V _l 5 89 GAAATTGTGTTGACGCAGTCTCCAGGCACC CTGTCTTTGTCTCCAGGGGAAAGAGCCACC CTCTCCTGCAGGGCCAGTCAGAGTGTTAGC GGCAACTACTTGGCCTGGTACCAGCAGAAA CCTGGCCAGGCTCCCAGGCTCCTCATCTAT GGTGCATCCAGCAGGGCCACTGGCATCCCA GACAGGTTCAGTGGCAGTGGGTCTGGGACA GACTTCACTCTCACCATCAGCAGACTGGAG CCTGAAGATTTTGCAGTGTATTATTGTCAG CAGTATGGTAGCGCACCGCTCACTTTCGGC GGAGGGACCAAGGTGGAAATCAAA 26H11 V _l 6 90 GAAATTGTGTTGACGCAGTCTCCAGGCACC CTGTCTTTGTCTCCAGGGGAAAGAGCCACC

101

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence CTCTCCTGCAGGGCCAGTCAGAGTGTTAGC GGCAACTACTTGGCCTGGTACCAGCAGAAA CCTGGCCAGGCTCCCAGGCTCCTCATCTAT GGTGCATCCAGCAGGGCCACTGGCATCCCA GACAGATTCAGTGGCAGTGGGTCTGGGACA GACTTCACTCTCACCATCAGCAGACTGGAG CCTGAAGATTTTGCAATGTATTATTGTCAG CAGTATGGTAGCTCACCGCTCACTTTCGGC GGAGGGTCCAAGGTGGAGATCAAA 12E4 V _l 7 91 GAAATTGTGTTGACGCAGTCTCCAGGCACC CTGTCTTTGTCTCCAGGGGAAAGAGCCACC CTCTCCTGCAGGGCCAGTCAGAATTTCGAC AGCAACTACTTAGCCTGGTACCAGCAGAAG CCTGGCCAGGCTCCCCGGCTCCTCATCTAT GGTGCATCCAGCAGGGCCACTGGCATCCCA GACAACTTCAGTGGCAGTGGGTCTGGGACA GACTTCACTCTCACCATCAGCAGACTGGAG CCTGAAGATTTTGCAATGTATTACTGTCAG CAGTATGGTAGTTCACCGCTCACTTTCGGC GGAGGGACCAAGGTGGAAATCAAA 12C11 V _l 8 92 GAAATTGTGTTGACGCAGTCTCCAGGCACC CTGTCTTTGTCTCCAGGGGAAAGGGCCACC CTCTCCTGCAGGGCCAGTCAGAATTTTGAC AGCAGCTCCTTAGCCTGGTACCAGCAGAAA CCTGGCCAGGCTCCCCGGCTCCTCATCTAT GGTGCATCCAGCAGGGCCACTGGCATCCCA GACAGGTTCAGGGGGACTGAC

102

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence CCTGAAGATTTTGCAATGTATTACTGTCAG CAGTGTGGTAGCTCACCGCTCACTTTCGGC GGAGGGACCAAGGTGGAAATCAAA 21H2 21B4 V _l 9 93 GAAATTGTGTTGACGCAGTCTCCAGGCACC CTGTCTTTGTCTCCAGGGGAAAGAGCCACC CTCTCCTGCAGGGCCAGTCAGAGTGTTAGC AGTACCTACTTAGCCTGGCACCAGCAGAAA CCTGGCCAGGGTCTTAGGCTCCTCATCTAT GGTGCATCCAGCAGGGCCACTGGCATCCCA GACAGGTTCAGTGGCAGTGGGTCTGGGACA GACTTCACTCTTACCATCAGCAGACTGGAG CCTGAAGATTTTGCAGTGTATTACTGTCAG CAGTATGGAAGCTCATTCACTTTCGGCGGA GGGACCAGGGTGGAGATCAAA 18B11.1 V _L 10 94 GATATTGTGATGACTCAGTCTCCACTCTCC CTGCCCGTCACCCCTGGAGAGCCGGCCTCC ATCTCCTGCAGGTCTAGTCAGAGCCTCCTG TATTATAATGGATTCACCTATTTGGATTGG TTCCTGCAGAAGCCAGGGCAGTCTCCACAT CTCCTGATCTATTTGGGTTCTAATCGGGCC TCCGGGGTCCCTGACAGGTTCAGTGGCAGT GTTTCAGGCACAGATTTTACACTGAAAATC AGCAGAGTGGAGGCTGAGGATGTTGGGGTT TATTATTGCATGCAGTCTCTGCAAACTCCA TTCACTTTCGGCCCTGGGACCAAAGTGGAT ATCAAA 18B11.2 V _L 11 95 GAAATAGTGATGACGCAGTCTCCAGCCACC CTGTCTGTGTCTCCAGGGGAAAGAGCCACC

103

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence CTCTCCTGCAGGGCCAGTCAGAGTGTTAAC AGCAACTTAGCCTGGTACCAGCAGAAACCT GGCCAGGCTCCCAGGCTCCTCATTTATGGT GTATCCACCAGGGCCACTGGTATCCCAGCC AGGTTCAGTGGCAGTGGGTCTGGGACAGAG TTCACTCTCACCATCCGCAGCCTGCAGTCT GAAGATTTTGCAGTTTATTACTGTCAGCAG TATAATAACTGGCCTCCGACGTTCGGCCAA GGGACCAAGGTGGAAATCAAA 2 0D4 V _l 12 96 GACATACAGCTGACCCAGTCTCCATCCTCC CTGTCTGCATCTATAGGAGACAGAGTCACC ATCACTTGCCGGGCAAGTCAGGACATTAGA TATGATTTAGGCTGGTATCAGCAGAAACCA GGGAAAGCCCCTAAGCGCCTGATCTATGCT GCATCCAGTTTGCAAAGTGGGGTCCCTTCA AGGTTCAGCGGCAGTGGATCTGGGACAGAA TTCACTCTCACAGTCAGCAGCCTGCAGCCT GAAGATTTTGCAACTTATTACTGTCTACAG CATAATAGTTACCCTCTCACTTTCGGCGGA GGGACCAAGGTGGAGATCGAA 46D11 V _l 13 97 GACATCCAGATGACCCAGTCTCCCTCTTCC GTGTCTGCATCTGTAGGAGACAGAGTCACC ATCACTTGTCGGGCGAGTCAGGGTATTAGC ATCTGGTTAGCCTGGTATCAGCAGAAACCT GGGAAAGCCCCTAAACTCCTGATCTATGCT GCATCCAGTTTGCAAAGTGGGGTCCCATCA AGGTTCAGCGGCAGTGGATCTGGGACAGAT TTCACTCTCACCATCAGCAGCCTGCAGCCT

104

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence GAAGATTTTGCAACTTACTATTGTCAACAG GCTAACGATTTCCCGATCACCTTCGGCCAA GGGACACGACTGGAGATTAAA 40D2 V _l 14 98 GATTTTGTGATGACCCAGACTCCACTCTCT CTGTCCGTCACCCCTGGACAGCCGGCCTCC ATCTCCTGCAAGTCTAGTCAGAGCCTCCTA CAGAGTGATGGAAAGACCTATTTGTATTGG TACCTGCAGAAGCCAGGCCAGCCTCCACAT CTCCTGATCTATGAAGTTTCCAACCGATTC TCTGGAGTGCCAGATAGGTTCAGTGGCAGC GGGTCAGGGACAGATTTCACACTGAAAATC AGCCGGGTGGAGGCTGAGGATGTTGGGGTT TATTACTGCATGCAAAGTATACAGCTTCCT CGGACGTTCGGCCAAGGGACCAAGGTGGAA ATCAAA 37D3 V _l 15 99 GATATTGTGATGACTCAGTCTCCACTCTCC CTGCCCGTCACCCCTGGAGAGCCGGCCTCC ATCTCCTGCAGGTCTAGTCAGAGCCTCCTG CATAGTAATGGATACAACTTTTTGGATTGG TACCTACAGAAGCCAGGGCAGTCTCCACAG CTCCTGATCTATTTGGGTTCTGATCGGGCC TCCGGGGTCCCTGACAGGTTCAGTGGCAGT GGATCAGGCACAGAGTTTACACTGAAAATC AGCAGAGTGGAGGCTGAGGATGTTGGGCTT TATTACTGCATGCAAGCTCTACAAACTCCG TGCAGTTTTGGCCAGGGGACCAAGCTGGAG ATCAAA 39F7 V _l 16 100 GAAATTGTGTTGACGCAGTCTCCAGGCACC

105

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence CTGTCTTTGTCTCCAGGGGAAAGAGCCACC CTCTCCTGCAGGGCCAGTCAGAGTGTTAGT AGCACCTATTTAGCCTGGTACCAGCAGAAA CCTGGCCAGGCTCCCAGGCTCCTCATCTAT GGTGCATCCAGCAGGGCCACTGGCATCCCA GACAGGTTCAGTGGCAGTGGGTCTGGGACA GACTTCACTCTCACCATCAGCAGACTGGAG CCTGAAGATTTTGCAGTTTATTACTGTCAG CAGTCTGGTAGCTCACCGCTCACTTTCGGC GGAGGGACCGAGGTGGAGATCAAA 39F11 V _l 17 101 GAAATTGTGTTGACGCAGTCTCCAGGCACC CTGTCTTTGTCTCCAGGGGAAAGAGCCACC CTCTCCTGCAGGGCCAGTCAGAGTGTTAGC AGCACCTACTTAGCCTGGTACCAGCAGAAA CCTGGCCAGGCTCCCAGTCTCCTCATCTAT GGTGCATCCAGCAGGGCCACTGGCATCCCA GACAGGTTCAGTGGCAGTGGGTCTGGGACA GACTTCACTCTCACCATCAGCAGACTGGAG CCTGAGGATTTTGCAGTGTATTACTGTCAG CAGTCTGGTAGCTCACCTCTCACTTTCGGC GGAGGGACCAAGGTGGAGATCAAA 3 0G5 V _l 18 102 GAAATTGTGTTGACGCAGTCTCCAGGCACC CTGTCTTTGTCTCCAGGGGAAAGAGCCACC CTCTCCTGCAGGGCCAGTCAGAGTGTTAGC AGCACCTACTTAGCCTGGTACCAGCAGAAA CCTGGCCAGGCTCCCAGGCTCCTCATCTAT GGTGCATCCTTCAGGGGGGGGGGGA

106

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence GACTTCACTCTCACCATCAGCAGACTGGAG CCTGAGGATTTTGCAGTGTATTACTGTCAG CAGTCTGGTAGCTCACCTCTCACTTTCGGC GGAGGGACCAAGGTGGAGATCAAAA

Table 2D - Coding sequence for variable heavy chains (V _H ) of antibody.

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence 17C3 V _H 1 103 CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCGTCTCTGGGTTCTCACTCAGC AATGCTAGAATGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACGAAAAATCC TACAGCACATCTCTGAAGAGCAGGCTCACC ATCTCCAAGGACACCTCCAAAAGCCAGGTG GTCCTTACCATGACCAACATGGACCCTGTG GACACAGCCACATATTACTGTGCACGGATA TTATTACTGGGAGCTTACTACTACTACGGT ATGGACGTCTGGGGCCAAGGGACCACGGTC ACCGTCTCCTCA 22H5 V _h 2 104 CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCGTCTCTGGGTTCTCACTCAGC AATGCTAGAATGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACCAGA

107

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence ATCTCCAAGGACACCTCCAAAAGCCAGGTG GTCCTTACCATGACCAACATGGACCCTGTG GACACAGCCACATATTACTGTGCACGGATA TTATTAGTGGGAGCTTACTACTACTGCGGT ATGGACGTCTGGGGCCAAGGGCCA 16H7 V _h 3 105 CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCGTCTCTGGGTTCTCACTCAAC AATGCTAGAATGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACGAAAAATCC TACAGCACATCTCTGAAGAGCAGGCTCACC ATCTCCAAGGACACCTCCAAAAGCCAGGTG GTCCTAATTATGACCAACATGGACCCTGTG GACACAGCCACATATTACTGTGCACGGTCA GTAGTAACTGGCGGCTACTACTACGACGGT ATGGACGTCTGGGGCCAAGGGACCACGGTC ACCGTCTCCTCA 24H11 V _h 4 106 CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCGTCTCTGGGTTCTCACTCAGC AATGCTAGAATGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACGAAAAATCC TACAGCACATCTCTGAAGAACAGGCTCACC ATCTCCAAGGACACCTCCAAAAGCCAGGTG GTCCTTATTATGACCAACATGGACCCTGTG

108

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence GACACAGCCACATATTACTGTGCACGGTCA GTAGTGACTGGCGGCTACTACTACGACGGT ATGGACGTCTGGGGCCAAGGGACCACGGTC ACCGTCTCCTCA 18G1 V _h 5 107 GAGGTGCAGCTGTTGGAGTCTGGGGGAGGG TTGGTACAGCCGGGGGGGTCCCTGAGACTC TCCTGTGCAGCCTCTAGATTCACCTTTAGC ACCTATGCCATGAGCTGGGTCCGCCAGGCT CCAGGGAAGGGGCTGGAGTGGGTCTCAGGT ATTAGTGGTAGTGGTGTCAGCACACACTAC GCAGACTCCGTGAAGGGCCGGTTCACCATC TCCAGAGACAATTCCAAGAACACGCTGTAT CTGCAAATGAACAGCCTGAGAGCCGAGGAC ACGGCCGTATATTACTGTGCGAAATCCCTC ATTGTAGTAATAGTATATGCCCTTGACCAC TGGGGCCAGGGAACCCTGGTCACCGTCTCC TCA 17D8 V _h 6 108 GAGGTGCAGCTGTTGGAGTCTGGGGGAGGG TTGGTACAGCCGGGGGGGTACCTGAGACTC TCCTGTGCAGCCTCTGGATTCACGTTTAGT ACCTATGCCATGAGCTGGGTCCGCCAGGCT CCAGGGAAGGGACTGGAGTGGGTCTCAGCT ATCAGTGGTAGTGGTGTTAGCACATACTAC GCAGACTCCGTGAAGGGCCGGTTCACCATC TCCAGAGACAATTCCAAGAACACGCTGTAT CTGCAAATGAACAGCCTGAGAGCCGAGGAC ACGGCCGTATATTACTGTGCGAAATCCCTT ATTGTAGTAATGGTGTATGTCCTTGACTAC

109

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence TGGGGCCAGGGAACCCTGGTCACCGTCTCC TCA 26H11 V _h 7 109 GAGGTGCAGCTGTTGGAGTCTGGGGGAGGC TTGGTACAGCCGGGGGGGTACCTGAGACTC TCCTGTGCAGCCTCTGGATTCACGTTTAGC ACCTATGCCATGAGCTGGGTCCGCCAGGCT CCAGGGAAGGGACTGGAGTGGGTCTCAGCT ATTAGTGGCAGTGGTGTGAGCACAAACTAC GCAGACTCCGTGAAGGGCCGGTTCACCATC TCCAGAGACAATTCCAAGAACACGCTGTAT CTGCAAATGAACAGCCTGAGAGCCGAGGAC ACGGCCGTATATTACTGTGCGAAATCCCTT ATTGTAGTAATGGTGTATGTCCTTGACTAC TGGGGCCAGGGAACCCTGGTCACCGTCTCC TCA 12E4 12C11 V _h 8 110 GAGGTGCAGCTGTTGGAGTCTGGGGGAGGG TTGGTACAGCCGGGGGGGTCCCTGAGACTC TCCTGTGCAGCCTCTAGATTCACCTTTAGC ACCTATGCCATGAGCTGGGTCCGCCAGGCT CCAGGGAAGGGGCTGGAGTGGGTCTCAGGT ATTAGTGGTAGTGGTGTTAGCACATACTAC GCAGACTCCGTGAAGGGCCGGTTCACCATC TCCAGAGACAATTCCAAGAACACGCTGTAT CTGCAAATGAACAGCCTGAGAGCCGAGGAC ACGGCCGTATATTACTGTGCGAAATCCCTT ATTGTAGTAATAGTATATGCCCTTGACTAC TGGGGCCAGGGAACCCTGGTCACCGTCTCC TCA

110

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence 21H2 V _h 9 111 CAGGTGCAGCTGCAGGAGTCGGGCCCAGGA CTGGTGAAGCCTTCGGAGACCCTGTCCCTC ACCTGCACTGTCTCTGGTGGCTCCATCAGT AGTTACTACTGGAGCTGGATCCGGCAGCCC GCCGGGAAGGGACTGGAGTGGATTGGGCGT ATCTATACCAGTGGGAGCACCAACTACAAC CCCTCCCTCAAGAGTCGGGTCACCATGTCA AAAGACACGTCCAAGAACCAGTTCTCCCTG AAGCTGAGGTCTGTGACCGCCGCGGACACG GCCGTGTATTACTGTGCGAGAGATCCGGAC GGTGACTACTACTACTACGGTATGGACGTC TGGGGCCAAGGGACCTCGGTCACCGTCTCC TCA 21B4 V _h 10 112 CAGGTGCAGCTGCAGGAGTCGGGCCCAGGA CTGGTGAAGCCTTCGGAGACCCTGTCCCTC ACCTGCACTGTCTCTGGTGGCTCCATCAGT AGTTACTTCTGGAGCTGGATCCGGCAGCCC GCCGGGAAGGGACTGGAGTGGATTGGGCGT ATCTATACCAGTGGGAGCACCAACTACAAC CCCTCCCTCAAGAGTCGAGTCACCATGTCA ATAGACACGTCCAAGAACCAGTTCTCCCTG AAGCTGAGTTCTGTGACCGCCGCGGACACG GCCGTGTATTACTGTGCGAGAGATCCGGAC GGTGACTACTACTACTACGGTATGGACGTC TGGGGCCAAGGGACCACGGTCACCGTCTCC TCA 18B11.1 18B11.2 V _h 11 113 GAGGTGCAGCTGGTGGAGTCTGGGGGAGGC TTGGTAAAGCCTGGGGGGTCCCTTAGACTC

Ill

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence TCCTGTGCAGCCTCTGGATTCACTTTCAGT GACGCCTGGATGAGCTGGGTCCGCCAGGCT CCAGGGAAGGGGCTGGAGTGGGTTGGCCGT attaaaagcaaaactgatggtgggacaaca GACTACGCTGCACCCGTGAAAGGCAGATTC ACCATCTCAAGAGATGATTCAAAAAACACT CTGTATCTGCAAATGAACAGCCTGAAAACC GAGGACACAGCCGTGTATTTTTGTACCTCT ACGTATAGCAGTGGCTGGTACGTATGGGAC TACTACGGTATGGACGTCTGGGGCCAAGGG ACCACGGTCACCGTCTCCTCA 2 0D4 V _h 12 114 CAGGTCCAGCTGGTACAGTCTGGGGCTGAG GTGAAGAAGCCTGGGGCCTCAGTGAAGGTC TCCTGCAAGGTTTCGGGATACACCCTCACT GATTTATCCATGCACTGGGTGCGACAGGCT CCTGGAAAAGGGCTTGAGTGGATGGGAGGT TTTGATCCTGAAGATGGTGAAACAATCTAC GCACAGAAGTTCCAGGGCAGAATCACCATG ACCGAGGACACATCTACAGACACAGCCTAC ATGGAGCTGAGCAGCCTGAGATCTGAGGAC ACGGCCGTGTATTACTGTGCAAGTATTGTA GTAGTCCCAGCTGCTATACAGAGTTACTAC TACTACTACGGTATGGGCGTCTGGGGCCAA GGGACCACGGTCACCGTCTCCTCC 46D11 V _h 13 115 CAGGTCACCTTGAAGGAGGCTGGTCCTGTG TTGGTGAAACCCACAGAGACCCTCACGTTG ACCTGCACCGTCTCTGGGTTCTCACTCAGC AATGCTAGAATGGGTGTGAACTGGATCCGT

112

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACGAAAAATCC TACAGCACATCTCTGAAGAGCAGGCTCACC ATCTCCAAGGACACCTCCAAAAGCCAGGTG GTCCTTACCATGACCAACATGGACCCTGTG GACACAGCCACATATGATGATGATGAC 4 0D2 V _h 14 116 CAGGTGCAGCTGCAGGAGTCGGGCCCAGGA CTGGTGAAGCCTTCACAGACCCTGTCCCTC ACCTGCACTGTCTCTGGTGGCTCCATCAGC AGTGGTGGTTACAACTGGAGCTGGATCCGC CAGCACCCAGGGAAGGGCCTGGAGTGGATT GGGAACATCTATTACAGTGGGAGCACCTAC TACAACCCGTCCCTCAAGAGTCGAGTTACC ATATCAGTAGACACGTCTAAGAACCAGTTC TCCCTGAAGCTGAGATCTGTGACTGCCGCG GACACGGCCGTGTATTACTGTGCGAGAGAG AATATTGTAGTAATACCAGCTGCTATATTC GCGGGTTGGTTCGACCCCTGGGGCCAGGGA ACCCTGGTCACCGTCTCCTCA 37D3 V _h 15 117 GAGGTGCACCTGGTGGAGTCTGGGGAGAGC TTGGCAAAGCCTGGGGGGTCCCTTAGACTC TCCTGTGCAGCCTCTGGATTCACTTTCAGA AACGCCTGGATGAGCTGGGTCCGCCGGGGGGGGGGGA

113

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence GACTACGCTGCACCCGTGAAAGGCAGATTC ACCATCTCGAGAGATGATTCAAAAAACACG CTGTATCTGCAAATGAACAGCCTGAAAACC GAGGACACAGCCGAGTATTACTGTATCACA GATCGGGTGCTAAGCTACTACGCTATGGCC GTCTGGGGCCAGGGGCCA 39F7 V _h 16 118 CAGGTGCAGCTGGTGGAGTCTGGGGGAGGC GTGGTCCAGCCTGGGAGGTCCCTGAGACTC TCCTGTGCAGCGTCTGGATTCACCTTCAGT AACTATGGCATTCACTGGGTCCGCCAGGCT CCAGGCAAGGGGCTGGAGTGGGTGGCAGTT ATATGGTATGATGGAAGTATTAAATACTAT GCAGACTCCGTGAAGGGCCGATTCACCATC TCCAGAGACAATTCCAAGAACACGCTGTAT CTGCAAATGAACAGCCTGAGAGCCGAGGAC ACGGCTGTGTATTACTGTGCGAGAGATAGG GCAGCAGCTGGTCTCCACTACTACTACGGT ATGGACGTCTGGGGCCAAGGGACCACGGTC ACCGTCTCCTCA 39F11 V _h 17 119 CAGGTGCAGCTGGTGGAGTCTGGGGGAGGC GTGGTCCAGCCTGGGAGGTCCCTGAGACTC TCCTGTGCAGCGTCTGGATTCACCTTCAGT AGCTATGGCATCCACTGGGTCCGCCAGGCT CCAGGCAAGGGGCTGGAATGGGTGGCAGTT ATATGGTATGATGGAAGTGATAAATACTAT GCAGACTCCGTGAAGGGCCGATTCACA

114

Contained in the clone Designation ID. IN SEQ. N °: Coding sequence CTACAAATGAACAGCCTGAGAGCCGAGGAC ACGGCTGTGTATTACTGTGCGAGAGATAGG GCAGCAGCTGGTCTCCACTATTATTACGGT ATGGACGTCTGGGGCCAAGGGACCACGGTC ACCGTCTCCTCA 39G5 V _h 18 120 CAGGTGCAGCTGGTGGAGTCTGGGGGAGGC GTGGTCCAGCCTGGGAGGTCCCTGAGACTC TCCTGTGCAGTGTCTGGATTCACCTTCAGT AGCTATGGCATCCACTGGGTCCGCCAGGCT CCAGGCAAGGGGCTGGAATGGGTGGCAGTT ATATGGTATGATGGAAGTGATAAATACTAT GGAGACTCCGTGAAGGGCCGATTCACCATC TCCAGAGACAATTCCAAGAACACGCTGTAT CTACAAATGAACAGCCTGAGAGCCGAGGAC ACGGCTGTGTATTACTGTGCGAGAGATAGG GCAGCAGCTGGTCTCCACTATTATTACGGT ATGGACGTCTGGGGCCAAGGGACCACGGTC ACCGTCTCCTCA

Each of the heavy chain variable regions listed in Table 2B can be combined with any of the light chain variable regions shown in Table 2A to form an antigen binding protein. Examples of such combinations include V _H 1 combined with any of V _L 1, V _l 2, V _l 3, V _L 4, V _L 5, V _L 6, V _L 7, V _L 8, V _L 9, V _L 10, V _L 11,

V _l 12, V _l 13, V _l 14, V _l 15, V _l 16, V _l 17 or V _l 18; V _h 2 combined with any of V _L 1, V _L 2, V _L 3, V _L 4, V _L 5, V _L 6, V _L 7, V _L 8, V

V _L 10, V _L 11, V _l 12, V _l 13, V _l 14, V _L 15, V _L 16, V _L 17 or V _L 18; V combined with any one of V _L 1, V _L 2, V _L 3, V _L 4, V _L 5, V

115

V _l 7, V _l 8, V _l 9, V _l 10, V _l 11, V _l 12, V _l 13, V _L 14, V _L 15, V _L 16, V _L 17 or V _l 18; and so on.

In some cases, the antigen-binding protein includes at least one heavy chain variable region and / or a light chain variable region from those listed in Tables 2A and 2B. In some cases, the antigen binding protein includes at least two variable regions of the heavy chain and / or variable regions of the light chain other than those listed in Table 2B. An example of such an antigen binding protein comprises (a) a V _H 1, and (b) one of V _H 2, V _h 3, V _H 4, V _H S, V _H 6, V _H 7, V _H 8, V _H 9, V _H 10, V _H 11, V _h 12, V _h 13, V _h 14, V _h 15, V _h 16, V _h 17 or V _h 18. Another example comprises (a) a V _H 2, and (b) a V _H 1, V _H 3, V _H 4, V _H S, V _H 6, V _h 7, V _h 8, V _h 9, V _h 10, V _h 11, V _h 12, V _h 13, V _H 14, V _H 15, V _H 16, V _H 17 or V _h 18. Again, another example comprises (a) a V _h 3, and ( b) one of V _H 1, V _H 2, V _H 4, V _H 5, V _H 6, V _H 7, V _H 8, V _H 9, V _H 10, V _H 11, V _h 12, V _h 13, V _h 14, V _h 15 V _h 16, V _h 17 or V _h 18 etc.

Again, another example of such an antigen binding protein comprises (a) a V _L 1, and (b) one of V _l 2, V _l 3, V _l 4, V _l 5, V _l 6, V _l 7, V _L 8, V _L 9, V _L 10, V _L 11, V _L 12, V _L 13, V _l 14, V _l 15, V _l 16, V _l 17 or V _l 18. Again, another example such an antigen binding protein comprises (a) a V _L 2, and (b) one of V _L 1, V _L 3, V _L 4, V _L 5, V _L 6, V _L 7, V _L 8, V _l 9, V _l 10, V _l 11 or V _l 12. Again, another example of such an antigen binding protein comprises (a) a V _L 3, and (b) one of V _L 1, V _L 2, V _L 4, V _L 5, V _L 6, V _L 7, V _L 8, V _L 9, V _L 10, V _L 11, V _l 12, V _l 13, V _l 14, V _L 15, V _L 16, V _L 17 or V _L 18 etc.

The various combinations of variable regions of the heavy chain can be combined with any of the various combinations of variable regions of the light chain.

116

In other embodiments, an antigen-binding protein comprises two identical light chain variable regions and / or two identical heavy chain variable regions. As an example, the antigen binding protein can be an antibody or immunologically functional fragment thereof that includes two variable regions of the light chain and two variable regions of the heavy chain in combinations of pairs of variable regions of the light chain and pairs of variable regions of the chain as listed in Tables 2A and 2B.

Some antigen binding proteins that are provided comprise a variable domain of the heavy chain that comprises an amino acid sequence that differs from the sequence of a variable domain of the selected heavy chain of V _H 1, V _H 2, V _H 3, V _H 4 , V _H 5, V _H 6, V _H 7, V _H 8, V _H 9, V _H 10, V _h 11, V _h 12, V _h 13, V _h 14, V _H 15, V _H 16, V _H 17 and V _H 18 in just 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 amino acid residues, where each of these sequence differences is independently a deletion, insertion or substitution of an amino acid, with deletions, insertions and / or substitutions resulting in a maximum of 15 amino acid changes in relation to the variable domain sequences mentioned above. The variable region of the heavy chain, in some antigen binding proteins, comprises a sequence of amino acids that has at least 70%, 75%, 80%, 85%, 90%, 95%, 97% or 99% identity sequence for the amino acid sequences of the variable region of the heavy chain of V _H 1, V _H 2, V _H 3, V _H 4, V _H S, V _H 6, V _H 7, V _H 8, V _H 9, V _H 10, V _H 11, V _H 12, V _h 13, V _h 14, V _H 15, V _H 16, V _H 17 and V _H 18.

117

Certain antigen binding proteins comprise a light chain variable domain that comprises an amino acid sequence that differs from the sequence of a selected light chain variable domain from V _L 1, V _L 2, V _l 3, V _l 4, V _L 5, V _l 6, V _l 7, V _l 8, V _L 9, V _L 10, V _L 11, V _L 12, V _L 13, V _l 14, V _l 15, V _l 16, V _l 17 e V _L 18 in just 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 amino acid residues, where each of these sequence differences is independently a deletion , insertion or substitution of an amino acid, with deletions, insertions and / or substitutions resulting in a maximum of 15 amino acid changes in relation to the variable domain sequences mentioned above. The light chain variable region on some antigen-binding proteins comprises an amino acid sequence that has at least 70%, 75%, 80%, 85%, 90%, 95%, 97% or 99% sequence identity for the amino acid sequences of the V _L 1, V _L 2, V _L 3, V _L 4, V _L 5, V _L 6, V _L 7, V _L 8, V _L 9, V _L 10 light chain variable region , V _L 11, V _l 12, V _l 13, V _l 14, V _L 15, V _L 16, V _L 17 or V _L 18.

In additional cases, antigen binding proteins comprise the following light and heavy chain variable domain pairings: V _L 1 with V _H 1, V _l 2 with V _h 2, V _l 2 with V _H 3, V _L 3 with V _H 4, V _L 4 with V _H 5, V _L 5 with

V _H 6, V _l 6 with V _h 7, V _L 7 with V _H 8, V _L 8 with V _H 8, V _L 9 with V _H 9, V with V _H 10, V _l 10 with V _H 11, V _L 11 with V _H 11, V _L 12 with V _H 12, V with V _h 13, V _l 14 with V _H 14, V _L 15 with V _H 15, V _L 16 with V _H 16, V with V _h 17, and V _l 18 with V _H 18. In some cases, the antigen-binding proteins in the above pairings may comprise sequences of amino acids that have 70%, 75%, 80%, 85%,

118

90%, 95%, 96%, 97%, 98% or 99% sequence identity with the specified variable domains.

Still other antigen binding proteins, for example, immunologically functional antibodies or fragments, include variant forms of a variant heavy chain and a variant light chain as described above.

Antigen-binding protein CDRs

In various embodiments, the antigen binding proteins disclosed herein can comprise polypeptides in which one or more CDRs are grafted, inserted and / or joined. An antigen-binding protein can have 1, 2, 3, 4, 5 or 6 CDRs. Thus, an antigen binding protein can have, for example, a heavy chain CDR1 (CDRH1) and / or a heavy chain CDR2 (CDRH2) and / or a heavy chain CDR3 (CDRH3) and / or a CDR1 light chain (CDRL1) and / or a light chain CDR2 (CDRL2) and / or a light chain CDR3 (CDRL3). Some antigen-binding proteins include both a CDRH3 and a CDRL3. Specific heavy and light chain CDRs are identified in Tables 3A and 3B, respectively, and in Table 6C, below.

The complementarity determining regions (CDRs) and framework regions (FRs) of a certain antibody can be identified using the system described by Kabat et al., In Sequences of Proteins of Immunological Interest, 5th edition, US Dept. of Health and Human Services, PHS, NIH, NIH Publication No. 91-3.242, 1991. As desired, CDRs can also be redefined according to an alternative nomenclature scheme, for example, that of Chothia (see Chothia and Lesk, 1987, J. Mol. Biol.

119

196: 901-917; Chothia et al., 1989, Nature 342: 878-883 or Honegger and Pluckthun, 2001, J. Mol. Biol. 309: 657-670).

Certain antibodies that are disclosed herein comprise one or more amino acid sequences that are identical or have substantial sequence identity for the amino acid sequences of one or more of the CDRs shown in Table 3A (CDRHs) and Table 3B (CDRLs) and Table 6C, infra.

Table 3A - Exemplary CDRH sequences.

Clone ID. SEQ. N °: Contained in the reference Designation Sequence 2 0D4 121 V _h 12 CDRH1-1 DLSMH 17C3 22H5 16H7 24H11 122 V _H 1 V _h 2 V _h 3 V _h 4 CDRH1-2 NARMGVS 18B11.1 18B11.2 123 V _h 11 V _h 11 CDRH1-3 DAWMS 18G1 V _h 5 12C11 V _h 8 12E4 124 V _h 8 CDRH1-4 TYAMS 17D8 V _h 6 26H11 V _h 7 21B4 125 V _h 10 CDRH1-5 SYFWS 46D11 126 V _h 13 CDRH1-6 NARMGVN 37D3 127 V _h 15 CDRH1-7 NAWMS 39F11 3 9G5 128 V _h 17 V _h 18 CDRH1-8 SYGIH 39F7 129 V _h 16 CDRH1-9 NYGIH

120

Clone ID. SEQ. N °: Contained in the reference Designation Sequence 40D2 130 V _h 14 CDRHl-10 SGGYNWS 21H2 131 V _h 9 CDRH1-11 SYYWS 20D4 132 V _h 12 CDRH2-I GFDPEDGETIYAQKFQG 17C3 22H5 16H7 46D11 133 V _h 1 V _h 2 V _h 3 V _h 13 CDRH2-2 HIFSNDEKSYSTSLKS 24H11 134 V _h 4 CDRH2-3 HIFSNDEKSYSTSLKN 18B11.1 18B11.2 37D3 135 v _h ii V _H ll V _h 15 CDRH2-4 RIKSKTDGGTTDYAAPVKG 18G1 136 V _h 5 CDRH2-5 GISGSGVSTHYADSVKG 12C11 12E4 137 V _h 8 V _h 8 CDRH2-6 GISGSGVSTYYADSVKG 17D8 138 V _h 6 CDRH2-7 AISGSGVSTYYADSVKG 26H11 139 V _h 7 CDRH2- 8 AISGSGVSTNYADSVKG 21B4 21H2 140 V _h 10 V _h 9 CDRH2- 9 RIYTSGSTNYNPSLKS 39F11 141 V _h 17 CDRH2-10 VIWYDGSDKYYADSVKG 39F7 142 V _h 16 CDRH2-11 VIWYDGSIKYYADSVKG 3 9G5 143 V _h 18 CDRH2-12 VIWYDGSDKYYGDSVKG 4 0D2 144 V _h 14 CDRH2-13 NIYYSGSTYYNPSLKS 2 0D4 145 V _h 12 CDRH3-1 IVWPAAIQSYYYYYGMGV 17C3 146 V _h 1 CDRH3-2 ILLLGAYYYYGMDV 22H5 147 V _h 2 CDRH3-3 ILLVGAYYYCGMDV

121

Clone ID. SEQ. N °: Contained in the reference Designation Sequence 16H7 24H11 148 V _h 3 V _h 4 CDRH3-4 SWTGGYYYDGMDV 18B11.1 18B11.2 149 v _h ii V _H 11 CDRH3-5 TYS SGWYVWDYYGMDV 18G1 150 V _h 5 CDRH3- 6 SLIWIVYALDH 12C11 12E4 151 V _h 8 V _h 8 CDRH3- 7 SLIWIVYALDY 17D8 26H11 152 V _h 6 V _h 7 CDRH3- 8 SLIVVMVYVLDY 21B4 21H2 153 V _H 10 V _H 9 CDRH3- 9 DPDGDYYYYGMDV 46D11 154 V _h 13 CDRH3-10 VRIAGDYYYYGMDV 3 7D3 155 V _h 15 CDRH3-11 DRVLSYYAMAV 39F11 V _h 17 39F7 156 V _h 16 CDRH3-12 DRAAAGLHYYYGMDV 39G5 V _h 18 4 0D2 157 V _h 14 CDRH3-13 ENIWIPAAIFAGWFDP

Table 3B - Exemplary CDRL sequences.

Clone ID. SEQ. N °: Contained in the reference Designation Sequence 20D4 158 V _l 12 CDRL1-1 RASQDIRYDLG 18B11.1 159 V _L 10 CDRL1-2 RSSQSLLYYNGFTYLD 12C11 160 V _l 8 CDRL1-3 RASQNFDSSSLA 18G1 161 V _l 4 CDRL1-4 RASQNFDSSYLA 17D8 26H11 162 V _l 5 V _l 6 CDRL1-5 RASQSVSGNYLA

122

Clone ID. SEQ. N °: Contained in the reference Designation Sequence 21B4 V _l 9 21H2 V _l 9 39F7 163 V _l 16 CDRL1-6 RASQSVSSTYLA 39F11 V _l 17 3 9G5 V _l 18 12E4 164 V _l 7 CDRL1-7 RASQNFDSNYLA 18B11.2 165 V _L 11 CDRL1-8 RASQSVNSNLA 16H7 24H11 166 V _l 3 V _l 3 CDRL1-9 GGNNIGSESVH 22H5 17C3 167 V _l 2 V _L 1 CDRL1-10 GGNNIGSQSVH 46D11 168 V _l 13 CDRL1-11 RASQGISIWLA 4 0D2 169 V _l 14 CDRL1-12 KSSQSLLQSDGKTYLY 3 7D3 170 V _l 15 CDRL1-13 RSSQSLLHSNGYNFLD 2 0D4 46D11 171 V _l 12 V _l 13 CDRL2-1 AASSLQS 18B11.1 172 V _L 10 CDRL2-2 LGSNRAS 12C11 V _l 8 17D8 V _l 5 21B4 V _l 9 21H2 26H11 173 V _l 9 V _l 6 CDRL2-3 GASSRAT 12E4 V _l 7 39F7 V _l 16 39F11 V _l 17 18G1 174 V _l 4 CDRL2-4 GTSSRAT 18B11.2 175 V _L 11 CDRL2-5 GVSTRAT

123

Clone ID. SEQ. N °: Contained in the reference Designation Sequence 16H7 V _l 3 24H11 22H5 176 V _l 3 V _l 2 CDRL2-6 DDSDRPS 17C3 V _L 1 40D2 177 V _l 14 CDRL2- 7 EVSNRFS 37D3 178 V _l 15 CDRL2- 8 LGSDRAS 39G5 179 V _l 18 CDRL2- 9 GASFRAT 2 0D4 180 V _l 12 CDRL3-1 LQHNSYPLT 18B11.1 181 V _L 10 CDRL3-2 MQSLQTPFT 12C11 182 V _l 8 CDRL3-3 QQCGSSPLT 18G1 183 V _l 4 CDRL3-4 QQYGGSPLT 17D8 184 V _l 5 CDRL3-5 QQYGSAPLT 21B4 21H2 185 V _l 9 V _l 9 CDRL3- 6 QQYGSSFT 26H11 12E4 186 V _l 6 V _l 7 CDRL3-7 QQYGSSPLT 18B11.2 187 V _L 11 CDRL3- 8 QQYNNWPPT 16H7 24H11 188 V _l 3 V _l 3 CDRL3- 9 QVWDGNSDHW 22H5 189 V _l 2 CDRL3-10 QVWDNTSDHW 17C3 190 V _L 1 CDRL3-11 QVWDSSSDHW 46D11 191 V _l 13 CDRL3-12 QQANDFPIT 4 0D2 192 V _l 14 CDRL3-13 MQSIQLPRT 37D3 193 V _l 15 CDRL3-14 MQALQTPCS 39F7 V _l 16 39F11 194 V _l 17 CDRL3-15 QQSGSSPLT 3 9G5 V _l 18

Table 3C - Coding sequences for CDRHs

124

Clone ID. SEQ. N °: Contained in the referendum Designation Sequence 2 0D4 195 V _h 12 CDRH1-1 GATTTATCCATGCAC 17C3 22H5 16H7 24H11 196 V _h 1 V _h 2 V _h 3 V _h 4 CDRH1-2 AATGCTAGAATGGGTGTGAG Ç 18B11.1 18B11.2 197 V _H ll V _h 11 CDRH1-3 GACGCCTGGATGAGC 18G1 12C11 12E4 17D8 26H11 198 V _h 5 V _h 8 V _h 8 V _h 6 V _h 7 CDRH1-4 ACCTATGCCATGAGC 21B4 21H2 199 V _h 10 V _h 9 CDRH1-5 AGTTACTTCTGGAGC 46D11 200 V _h 13 CDRH1-6 AATGCTAGAATGGGTGTGAA Ç 37D3 201 V _h 15 CDRH1-7 AACGCCTGGATGAGC 39F11 3 9G5 202 V _h 17 V _h 18 CDRH1-8 AGCTATGGCATCCAC 39F7 203 V _h 16 CDRH1-9 AACTATGGCATTCAC 4 0D2 204 V _h 14 CDRH1-10 AGTGGTGGTTACAACTGGAG Ç 2 0D4 205 V _h 12 CDRH2-1 GGTTTTGATCCTGAAGATGG TGAAACAATCTACGCACAGA AGTTCCAGGGC

125

Clone ID. SEQ. N °: Contained in the reference Designation Sequence 17C3 22H5 16H7 46D11 206 V _H 1 V _h 2 V _h 3 V _h 13 CDRH2-2 CACATTTTTTCGAATGACGA AAAATCCTACAGCACATCTC TGAAGAGC 24H11 207 V _h 4 CDRH2-3 CACATTTTTTCGAATGACGA AAAATCCTACAGCACATCTC TGAAGAAC 18B11.1 18B11.2 37D3 208 V _h 11 V _h 11 V _h 15 CDRH2-4 CGTATTAAAAGCAAAACTGA TGGTGGGACAACAGACTACG CTGCACCCGTGAAAGGC 18G1 209 V _h 5 CDRH2-5 GGTATTAGTGGTAGTGGTGT CAGCACACACTACGCAGACT CCGTGAAGGGC 12C11 12E4 210 V _h 8 V _h 8 CDRH2- 6 GGTATTAGTGGTAGTGGTGT TAGCACATACTACGCAGACT CCGTGAAGGGC 17D8 211 V _h 6 CDRH2-7 GCTATCAGTGGTAGTGGTGT TAGCACATACTACGCAGACT CCGTGAAGGGC 26H11 212 V _h 7 CDRH2- 8 GCTATTAGTGGCAGTGGTGT GAGCACAAACTACGCAGACT CCGTGAAGGGC

126

Clone ID. SEQ. N °: Contained in the reference Designation Sequence 21B4 21H2 213 V _H 10 V _H 9 CDRH2- 9 CGTATCTATACCAGTGGGAG CACCAACTACAACCCCTCCC TCAAGAGT 39F11 214 V _h 17 CDRH2-10 GTTATATGGTATGATGGAAG TGATAAATACTATGCAGACT CCGTGAAGGGC 39F7 215 V _h 16 CDRH2-11 GTTATATGGTATGATGGAAG TATTAAATACTATGCAGACT CCGTGAAGGGC 3 9G5 216 V _h 18 CDRH2-12 GTTATATGGTATGATGGAAG TGATAAATACTATGGAGACT CCGTGAAGGGC 40D2 217 V _h 14 CDRH2-13 AACATCTATTACAGTGGGAG CACCTACTACAACCCGTCCC TCAAGAGT 20D4 218 V _h 12 CDRH3-1 ATTGTAGTAGTCCCAGCTGC TATACAGAGTTACTACTACT ACTACGGTATGGGCGTC 17C3 219 V _h 1 CDRH3-2 ATATTATTACTGGGAGCTTA CTACTACTACGGTATGGACG TC 22H5 220 V _h 2 CDRH3-3 ATATTATTAGTGGGAGCTTA CTACTACTGCGGTATGGACG TC 16H7 221 V _h 3 CDRH3-4 TCAGTAGTAACTGGCGGCTA

127

Clone ID. SEQ. N °: Contained in the reference Designation Sequence 24H11 V _h 4 CTACTACGACGGTATGGACG TC 18B11.1 18B11.2 222 V _h 11 v _h ii CDRH3-5 ACGTATAGCAGTGGCTGGTA CGTATGGGACTACTACGGTA TGGACGTC 18G1 223 V _h 5 CDRH3-6 TCCCTCATTGTAGTAATAGT ATATGCCCTTGACCAC 12C11 12E4 224 V _h 8 V _h 8 CDRH3-7 TCCCTTATTGTAGTAATAGT ATATGCCCTTGACTAC 17D8 26H11 225 V _h 6 V _h 7 CDRH3- 8 TCCCTTATTGTAGTAATGGT GTATGTCCTTGACTAC 21B4 21H2 226 V _h 10 V _h 9 CDRH3- 9 GATCCGGACGGTGACTACTA CTACTACGGTATGGACGTC 46D11 227 V _h 13 CDRH3-10 GTTCGTATAGCAGGTGATTA CTACTACTACTACGGTATGG ACGTC 3 7D3 228 V _h 15 CDRH3-11 GATCGGGTGCTAAGCTACTA CGCTATGGCCGTC 39F11 39F7 3 9G5 229 V _h 17 V _h 16 V _h 18 CDRH3-12 GATAGGGCAGCAGCTGGTCT CCACTATTATTACGGTATGG ACGTC 4 0D2 230 V _h 14 CDRH3-13 GAGAATATTGTAGTAATACC AGCTGCTATATTCGCGGGTT GGTTCGACCCC

Table 3D - Coding sequences for CDRLs.

128

Clone ID. IN SEQ. N °: Contained in the reference Designation Sequence 2 0D4 231 V _l 12 CDRL1-1 CGGGCAAGTCAGGACATTAGAT ATGATTTAGGC 18B11.1 232 V _L 10 CDRL1-2 AGGTCTAGTCAGAGCCTCCTGT ATTATAATGGATTCACCTATTT GGAT 12C11 233 V _l 8 CDRL1-3 AGGGCCAGTCAGAATTTTGACA GCAGCTCCTTAGCC 18G1 234 V _l 4 CDRL1-4 AGGGCCAGTCAGAATTTTGACA GCAGTTACTTAGCC 17D8 26H11 235 V _l 5 V _l 6 CDRL1-5 AGGGCCAGTCAGAGTGTTAGCG GCAACTACTTGGCC 21B4 21H2 39F7 39F11 3 9G5 236 V _l 9 V _l 9 V _l 16 V _l 17 V _l 18 CDRL1-6 AGGGCCAGTCAGAGTGTGAGCA GTACCTACTTAGCC 12E4 237 V _l 7 CDRL1-7 AGGGCCAGTCAGAATTTCGACA GCAACTACTTAGCC 18B11.2 238 V _L 11 CDRL1-8 AGGGCCAGTCAGAGTGTTAACA GCAACTTAGCC 16H7 24H11 239 V _l 3 V _l 3 CDRL1-9 GGGGGAAACAACATTGGAAGTG AAAGTGTGCAC 22H5 17C3 240 V _l 2 V _L 1 CDRL1-10 GGGGGAAACAACATTGGAAGTG AAAGTGTGCAC 46D11 241 V _l 13 CDRL1-11 CGGGCGAGTCAGGGTATTAGCA

129

Clone ID. SEQ. N °: Contained in the reference Designation Sequence TCTGGTTAGCC AAGTCTAGTCAGAGCCTCCTAC 4 0D2 242 V _L 14 CDRLl-12 AGAGTGATGGAAAGACCTATTT GTAT AGGTCTAGTCAGAGCCTCCTGC 3 7D3 243 V _l 15 CDRLl-13 ATAGTAATGGATACAACTTTTT GGAT 2 0D4 244 V _l 12 CDRL2-1 GCTGCATCCAGTTTGCAAAGT 46D11 V _l 13 18B11.1 245 V _L 10 CDRL2-2 TTGGGTTCTAATCGGGCCTCC 12C11 V _l 8 17D8 V _l 5 21B4 V _l 9 21H2 246 V _l 9 CDRL2- 3 GGTGCATCCAGCAGGGCCACT 26H11 V _l 6 12E4 V _l 7 39F7 V _l 16 39F11 V _l 17 18G1 247 V _l 4 CDRL2-4 GGTACATCCAGCAGGGCCACT 18B11.2 248 V _L ll CDRL2-5 GGTGTATCCACCAGGGCCACT 16H7 V _L 3 24H11 249 V _l 3 CDRL2- 6 GATGATAGCGACCGGCCCTCA 22H5 V _l 2 17C3 V _L l 40D2 250 V _l 14 CDRL2-7 GAAGTTTCCAACCGATTCTCT 37D3 251 V _l 15 CDRL2- 8 TTGGGTTCTGATCGGGCCTCC

130

Clone ID. SEQ. N °: Contained in the reference Designation Sequence 2 0D4 252 V _L 12 CDRL3-1 CTACAGCATAATAGTTACCCTC TCACT 18B11.1 253 V _L 10 CDRL3-2 ATGCAGTCTCTGCAAACTCCAT TCACT 12C11 254 V _l 8 CDRL3-3 CAGCAGTGTGGTAGCTCACCGC TCACT 18G1 255 V _l 4 CDRL3-4 CAGCAGTATGGTGGCTCACCGC TCACT 17D8 256 V _l 5 CDRL3- 5 CAGCAGTATGGTAGCGCACCGC TCACT 21B4 21H2 257 V _l 9 V _l 9 CDRL3-6 CAGCAGTATGGAAGTTCATTCA CT 26H11 12E4 258 V _l 6 V _l 7 CDRL3-7 CAGCAGTATGGTAGCTCACCGC TCACT 18B11.2 259 V _L 11 CDRL3- 8 CAGCAGTATAATAACTGGCCTC CGACG 16H7 24H11 260 V _l 3 V _l 3 CDRL3- 9 CAGGTGTGGGATGGTAATAGTG ATCATGTGGTA 22H5 261 V _l 2 CDRL3-10 CAGGTGTGGGATAATACTAGTG ATCATGTGGTA 17C3 262 V _L 1 CDRL3-11 CAGGTGTGGGATAGTAGTAGTG ATCATGTGGTA 46D11 263 V _L 13 CDRL3-12 CAACAGGCTAACGATTTCCCGA TCACC 40D2 264 V _l 14 CDRL3-13 ATGCAAAGTATACAGCTTCCTC GGACG

131

Clone ID. IN SEQ. N °: Contained in the reference Designation Sequence 3 7D3 265 V _l 15 CDRL3-14 ATGCAAGCTCTACAAACTCCGT GCAGT 39F7 39F11 3 9G5 266 V _l 16 V _l 17 V _l 18 CDRL3-15 CAGCAGTCTGGTAGCTCACCTC TCACT

The structure and properties of CDRs within a naturally occurring antibody have been described, supra. Briefly, in a traditional antibody, CDRs are embedded within a framework in the variable region of the heavy and light chain where they constitute the regions responsible for antigen binding and recognition. A variable region comprises at least three heavy or light chain CDRs; see supra (Kabat et al., 1991, Sequences of Proteins of Immunological Interest, Public Health Service NIH, Bethesda, MD; see also Chothia and Desk, 1987, J. Mol. Biol. 196: 901-917; Chothia et al. , 1989, Nature 342: 877-883), within a framework region (referred to as framework regions 1-4, FR1, FR2, FR3 and FR4, by Kabat et al., 1991; see also Chothia and Lesk, 1987, supra) . The CDRs provided herein, however, can be used not only to define the antigen binding domain of a traditional antibody structure, but can be embedded in several other polypeptide structures, as described herein.

In one aspect, the CDRs provided are: (a) a CDRH selected from the group consisting of (i) a CDRH1 selected from the group consisting of the ID. SEQ. N °: 121

132

131; (ii) a CDRH2 selected from the group consisting of the ID. SEQ. No. 132-144; (iii) a CDRH3 selected from the group consisting of the ID. SEQ. No.: 145-157; and (iv) a CDRH of (i), (ii) and (iii) containing one or more amino acid substitutions, deletions or insertions of a maximum of five, four, three, two or one amino acid; (B) a CDRL selected from the group consisting of (i) a CDRL1 selected from the group consisting of the ID. SEQ. No. 158 158; (ii) a CDRL2 selected from the group consisting of the ID. SEQ. No.: 171-179; (iii) a CDRL3 selected from the group consisting of the ID. SEQ. No. 180-194; and (iv) a CDRL of (i), (ii) and (iii) containing one or more amino acid substitutions, deletions or insertions of a maximum of 1, 2, 3, 4 or 5 amino acids.

In another aspect, an antigen binding protein comprises 1, 2, 3, 4, 5 or 6 variant forms of the CDRs listed in Tables 3A and 3B and Table 6C, below, each having at least 80%, 85%, 90 %, 95%, 96%, 97%, 98% or 99% sequence identity for a CDR sequence listed in Tables 3A and 3B and Table 6C, below. Some antigen binding proteins comprise 1, 2, 3, 4, 5 or 6 of the CDRs listed in Tables 3A and 3B and Table 6C, below, each differing by a maximum of 1, 2, 3, 4 or 5 amino acids CDRs listed in these tables.

In yet another aspect, an antigen binding protein includes the following combinations of CDRL1, CDRL2 and CDRL3: IDS. SEQ. ^Nos: 167, 176 and 190; IDS. SEQ. ^Nos: 167, 176 and 189, the IDS. SEQ. ^Nos: 166, 176 and 188; IDS. SEQ. ^Nos: 166, 176 and 188; IDS. SEQ. ^Nos: 161, 174 and 183; IDS. SEQ. ^Nos: 162, 173 and 184; IDS. SEQ. ^Nos: 162, 173

133

and 186; IDS. SEQ. ^Nos: 164, 173 and 186; IDS. SEQ. ^NOS: 160, 173 and 182; IDS . IN SEQ. ^Nos: 163, 173 and 185; IDS . IN SEQ. ^Nos: 163, 173 and 185, ; IDS. IN SEQ. ^Nos: 15 9 , 172 and 181; IDS. SEQ. ^Nos: 165, 175 and 187; IDS. SEQ. ^Nos: 158, 171 and 180 i; IDS. SEQ. ^Nos: 168, 171 and 191; IDS. SEQ. ^Nos: 169, 177 and 192; IDS . IN SEQ. ^Nos: 170, 178 and 193; IDS . IN SEQ. ^Nos: 163, 173 and 194, ; IDS. IN SEQ. ^Nos: 163 173 and 194; and IDS . SEQ. ^Nos: 163, 179 and 194 In one aspect additional, a protein binding

antigen includes the following associations of CDRH1, CDRH2 and CDRH3: IDS. SEQ. ^Nos: 122, 133 and 146; IDS. SEQ. ^Nos: 122, 133 and 147; IDS. SEQ. ^Nos: 122, 133 and 148; IDS. SEQ. ^Nos: 122, 134 and 148; IDS. SEQ. ^Nos: 124, 136 and 150;

IDS. SEQ. ^Nos: 124, 138 and 152; IDS. SEQ. ^Nos: 124, 139 and 152; IDS. SEQ. ^Nos: 124, 137 and 151; IDS. SEQ. ^Nos:

i

124, 137 and 151; IDS. SEQ. ^Nos: 131, 140 and 153; IDS. IN

SEQ. ^Nos: 125, 140 and 153; IDS. SEQ. ^Nos: 123, 135 and 149; IDS. SEQ. ^Nos: 123, 135 and 149; IDS. SEQ. ^Nos: 121, 132 and 145; IDS. SEQ. ^Nos: 126, 133 and 154; IDS. SEQ. ^Nos:

130, 144 and 157; IDS. SEQ. ^Nos: 127, 135 and 155; IDS. SEQ. ^Nos: 129, 142 and 156; IDS. SEQ. ^Nos: 128, 141 and 156; and IDS. SEQ. ^Nos: 128, 143 and 156.

In another aspect, an antigen-binding protein includes the following associations of CDRL1, CDRL2 and CDRL3 with

CDRH1, CDRH2 and CDRH3: IDS. SEQ. ^Nos: 167, 176 and 190; IDS. IN SEQ. N ^os : 167, 176 and 189, IDS. IN SEQ. ^Nos: 166, 176 and 188; IDS. IN SEQ. ^Nos: 166 176 and 188; IDS. SEQ. ^Nos: 161, 174 and 183; IDS. FROM SEQ . ^Nos: 162 173 and 184; IDS . IN SEQ. N ^os 162 , 173 and 186; IDS. IN SEQ. N ^{os 1} : 164 173 and 186; IDS. IN SEQ. N ^os : 160, 173 and 182; IDS. IN SEQ. ^Nos: 163, 173

134 ί

and 185; IDS. SEQ. ^Nos: 163, 173 and 185; IDS. SEQ. ^Nos: 159, 172 and 181; IDS. SEQ. ^Nos: 165, 175 and 187; IDS. SEQ. ^Nos: 158, 171 and 180; IDS. SEQ. ^Nos: 168, 171 and 191; IDS. SEQ. ^Nos: 169, 177 and 192; IDS. SEQ. ^Nos: 170, 178 and 193; IDS. SEQ. ^Nos: 163, 173 and 194; IDS. SEQ. ^Nos:

163, 173 and 194; IDS. SEQ. ^Nos: 163, 179 and 194 with IDS.

SEQ. ^Nos: 122, 133 and 146; IDS. SEQ. ^Nos: 122, 133 and

147; IDS. SEQ. ^Nos: 122, 133 and 148; IDS. SEQ. ^Nos: 122, 134 and 148; IDS. SEQ. ^Nos: 124, 136 and 150; IDS. IN

SEQ. ^Nos: 124, 138 and 152; IDS. SEQ. ^Nos: 124, 139 and 152; IDS. SEQ. ^Nos: 124, 137 and 151; IDS. SEQ. ^Nos: 124, 137 and 151; IDS. SEQ. ^Nos: 131, 140 and 153; IDS. SEQ. ^Nos: 125, 140 and 153; IDS. SEQ. ^Nos: 123, 135 and 149; IDS. SEQ. ^Nos: 123, 135 and 149; IDS. SEQ. ^Nos: 121, 132 and 145; IDS. SEQ. ^Nos: 126, 133 and 154; IDS. SEQ. ^Nos: 130, 144 and 157; IDS. SEQ. ^Nos: 127, 135 and 155; IDS. SEQ. ^Nos: 129, 142 and 156; IDS. SEQ. ^Nos: 128, 141 and 156; and IDS. IN

SEQ. ^Nos: 128, 143 and 156.

Consensus strings

In yet another aspect, the CDRs disclosed herein include consensus sequences derived from groups of related monoclonal antibodies. As described here, a consensus sequence refers to the amino acid sequences that have conserved amino acids common among several variable sequences and amino acids that vary within certain amino acid sequences. The CDR consensus strings provided include CDRs that correspond to each of CDRH1, CDRH2, CDRH3, CDRL1,

CDRL2 and CDRL3.

135

Consensus sequences were determined using standard CDR analyzes that correspond to the V _H and V _L of the revealed antibodies, some of which specifically bind to: (i) β-Klotho; (ii) FGFRlc, FGFR2c, 5 FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. The consensus sequences were determined by keeping the CDRs contiguous within the same sequence that corresponds to a V _H or V _L.

Light chain CDR3

Group 1 LQHNSYPLT (ID. IN SEQ. N °: 267) Group 2 MQSLQTPFT (ID. IN SEQ. N °: 268) 15 Group 3 QQYNNWPPT (ID. IN SEQ. N °: 269)

Group 4

MQSIQLPRT (ID. SEQ. N °: 270) Group 5 20 QQANDFPIT (ID. SEQ. N °: 271) Group 6 MQALQTPCS (ID. SEQ. N °: 272) Group 7 QVWD G N SDHW (ID. . SEQ. No. : 273) 25 QVWD N T SDHW (ID. . SEQ. No. : 274) QVWD S S SDHW (ID. . SEQ. No. : 275) QVWD Xi X ₂ SDHW (ID . SEQ. No. : 276) where Xi is G, S or N and X ₂ is s, T or N. Group 8 30 QQ C G S S P L T (SEQ ID. NO: 277)

136

QQ Y G G s P L T (ID. . SEQ. . N °: : 278) QQ Y G s THE P L T (ID. . SEQ. . N °: : 279) QQ Y G s s F T (ID. . SEQ. . N °: : 280) QQ Y G s s P L T (ID. . SEQ. . N °: : 281) QQ s G s s P L T (ID. . SEQ. . N °: : 282) QQ x ₃ G X ₄ χ ₅ x ₆ X ₇ T (ID. SEQ. N °: 283) in that X ₃ is C, Y or s, X ₄ is S or G, X ₅ is S or A, X ₆ is P

or F and X ₇ is L or absent.

Light chain CDR2

Group 1

AASSLQS (ID. IN SEQ. N °: 284) Group 2 GVSTRAT (ID. IN SEQ. N °: 285) Group 3 DDSDRPS (ID. IN SEQ. N °: 286) Group 4 EVSNRFS (ID. IN SEQ. N °: 287) Group 5

L G s N R A s (ID. IN SEQ. N °: 288) L G s D R A s (ID. IN SEQ. N °: 289) L G s X27 R A s (ID. IN SEQ. N °: 290) in what X ₂₇ is N or D.

Group 6

G THE s s RAT (ID. IN SEQ. N °: 291) G T s s RAT (ID. IN SEQ. N °: 292) G THE s F RAT (ID. IN SEQ. N °: 293) G χ ₈ s X28 RAT (ID. IN SEQ. N °: 294) in that X ₈ and the or T θ X28 is S or F.

Light chain CDR1

Group 1

137

RASQSVNSNLA (SEQ ID. NO: 295)

Group 2

RASQDIRYDLG (SEQ ID NO: 296)

Group 3

RASQGISIWLA (SEQ ID. NO: 297)

Group 4

KSSQSLLQSDGKTYLY (SEQ ID. NO: 298)

Group 5

RASQ N F D s s s THERE (ID. SEQ. No. : 299) RASQ N F D s s Y THERE (ID. SEQ. No. : 300) RASQ S V s G N Y THERE (ID. SEQ. No. : 301) RASQ S V s G T Y THERE (ID. SEQ. No. : 302) RASQ N F D s N Y THERE (ID. SEQ. No. : 303) RASQ X ₉ X10 X11 X12 X13 X14 THERE (ID. SEQ. No. : 304) in that X ₉ and the or S, X10 ê V or F X11 is D or s, X12 θ or S, X 13 is S, N or T, and X14 is S c JU Y.

Group 6

GGNNIGS AND SVH (ID . IN SEQ. N °: 305) GGNNIGS Q SVH (ID . IN SEQ. N °: 306) GGNNIGS X15 SVH (ID . IN SEQ. N °: 307) in that Xi5 and is or Q.

Group 7

RSSQSLL Y Y NG F T Y LD (ID. IN SEQ. No. 308) RSSQSLL H s NG Y N F LD (ID. IN SEQ. No. 309) RSSQSLL X29 X30 NG X31 X32 X33 LD (ID. IN SEQ. No. 310) in that X ₂ 9 is Y or H,  X30 is Y or S, X31 is F or Y , X32 is

or N and X ₃₃ is Y or F.

138

HEAVY CDR3

Group 1

IWVPAAIQSYYYYYGMGV (SEQ ID. N °: 311)

Group 2

DPDGDYYYYGMDV (SEQ ID. NO: 312)

Group 3

TYSSGWYVWDYYGMDV (SEQ ID. N °: 313)

Group 4

DRVLSYYAMAV (SEQ ID NO: 314)

Group 5

VRIAGDYYYYGMDV (SEQ ID. NO .: 315)

Group 6

ENIWIPAAIFAGWFDP (SEQ ID NO: 316)

Group 7

¹⁵ 4 DRAAAGLHYYYGMDV (ID Group 8 . FROM SEQ . N °: 317) I L L L G THE YYY Y GMDV (ID. IN SEQ N °: 318) I L L V G THE YYY Ç GMDV (ID. IN SEQ 20 N °: 319) V V T G G YYY D GMDV (ID. IN SEQ N °: 320) s V V T G G YYY D GMDV (ID. IN SEQ N °: 321) 25 X ₃₄ X16 X17 X18 G X19 YYY X20 GMDV (ID. IN SEQ N °: 322) in that X 34 is I, , V or S , X16 is L or v, x ₁₇ is L, T OR V Xis is L, V, G or T, X19 and the, G or absent and X ₂ the is Y, C or D. Group 9 30 SLIW I VY THE LD H ( ID. IN SEQ. N °: 323)

139

SLIW I VY THE LD Y (SEQ ID. N °: 324) SLIW M VY V LD Y (SEQ ID. N °: 325) SLIW X21 VY X22 LD X23 (SEQ ID. . No. : 326) in that X ₂ i Hey or M, X22 and the or V and X ₂₃ is H or Y.

HEAVY CDR2

Group 1

GFDPEDGETIYAQKFQG (SEQ ID. N °: 327)

Group 2

RIKSK T DGGTTDYAAPVKG (ID. IN SEQ. N °: 328) 10 RIKSK DGGTTDYAAPVKG (ID. IN SEQ. N °: 330) RIKSK X ₄ 2 DGGTTDYAAPVKG (ID. IN SEQ. N °: 483)

where X ₄₂ is T or absent.

Group 3

HIFSNDEKSYSTSLK S (SEQ ID NO: 331)

HIFSNDEKSYSTSLK N (SEQ ID. NO: 332)

HIFSNDEKSYSTSLK X ₂₄ (SEQ ID NO: 333) where X ₂₄ is S or N.

Group 4

G ISGSGVST H YADSVKG (ID. IN SEQ. N °: 334) 20 G ISGSGVST Y YADSVKG (ID. IN SEQ. N °: 335) THE ISGSGVST Y YADSVKG (ID. IN SEQ. N °: 336) THE ISGSGVST N YADSVKG (ID. IN SEQ. N °: 337) X25 ISGSGVST X ₂₆ YADSVKG (ID. IN SEQ. N °: 338)

where X ₂₅ is G or A and X ₂₆ is Η, Y or N.

Group 5

VIWYDGS D KYY THE DSVKG (ID . IN SEQ. N °: 339) VIWYDGS I KYY G DSVKG (ID . IN SEQ. N °: 340) VIWYDGS X35 KYY x ₃₆ DSVKG (ID . IN SEQ. N °: 341) in that X35 is D or I and X36 is A < DU G.

Group 6

140

N IY Y SGST Y YNPSLKS (ID. FROM SEQ . No. : 342) R IY T SGST Y YNPSLKS (ID. FROM SEQ . No. : 343) R IY T SGST N YNPSLKS (ID. FROM SEQ . No. : 329) X37 IY x ₃₈ SGST X ₄₁ YNPSLKS (SEQ ID. N °: 344) in that X37 is N or R, X ₃₈ is Y or T θ X41 θ Y or N.

HEAVY CDR1

Group 1

DLSMH (ID. Group SEQ. 2 N °: 345) 10 DAWNS (ID. IN SEQ. N °: 346) Group 3 TYAMS (ID. IN SEQ. N °: 347) Group 4 SYFWS (ID. IN SEQ. N °: 348) 15 Group 5 SGGYNWS (II d. : SEQ. N ⁰ : 349 Group 6

NARMGV S ( (ID. SEQ. N °: 350) NARMGV N ( : id. SEQ. N °: 351) 2 0 NARMGV X39 (ID . FROM SEQ . No. : 352)

where X ₃₉ is S or N. Group 7 s YGIH (ID. IN SEQ. N °: 353) N YGIH (ID. IN SEQ. N °: 354) 25 X ₄₀ YGIH (ID. IN SEQ. N °: 355) where X ₄₀ is S or N. In some cases, an antigen binding protein

comprises at least one heavy chain CDR1, CDR2 or CDR3 that has one of the consensus strings above. In some cases, an antigen-binding protein

141 comprises at least one light chain CDR1, CDR2 or CDR3 that has one of the consensus strings above. In other cases, the antigen binding protein comprises at least two heavy chain CDRs according to the consensus sequences above and / or at least two light chain CDRs according to the above consensus sequences. In still other cases, the antigen binding protein comprises at least three heavy chain CDRs according to the above consensus strings and / or at least three

Light chain CDRs according to the consensus strings above.

Exemplary antigen binding proteins

According to one aspect, an isolated antigen binding protein comprising (a) one or more heavy chain complementarity determining regions (CDRHs) / selected from the group consisting of: (i) a CDRH1 selected from the group consisting of in the ID. SEQ. No.: 121131; (ii) a CDRH2 selected from the group consisting of the ID. SEQ. No. 132-144; (iii) a CDRH3 selected from the group consisting of the ID. SEQ. No.: 145-157; and (iv) a CDRH of (i), (ii) and (iii) containing one or more amino acid substitutions, deletions or insertions of a maximum of 1, 2, 3, 4 or 5 amino acids; (b) one or more light chain complementarity determining regions (CDRLs) selected from the group consisting of: (i) a CDRL1 selected from the group consisting of the ID. SEQ. No. 158-170; (ii) a CDRL2 selected from the group consisting of the ID. SEQ. No.: 171-179; (iii) a CDRL3 selected from the group consisting of the ID. SEQ. No. 180-194; and (iv) a

CDRL of (i), (ii) (iii) containing one or more

142 amino acid substitutions, deletions or insertions of a maximum of five, four, three, four, two or one amino acid; or (c) one or more CDRHs of the heavy chain of (a) and one or more CDRLs of the light chain of (b).

In another modality, CDRHs have at least 70%,

75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of sequence identity with an amino acid sequence selected from the group consisting of the ID. SEQ. N °: 121-157 and / or CDRLs have at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 10 97%, 98% or 99% of sequence identity with a amino acid sequence selected from the group consisting of the ID. SEQ. No. 158-194. In an additional modality, the VH is selected from the group consisting of the ID. SEQ. N °: 121-157 and / or the VL is selected from the group consisting of the 15 ID. SEQ. No. 158-194.

According to one aspect, an isolated antigen-binding protein comprising (a) one or more variable heavy chains (VHs) selected from the group consisting of: (i) ID. SEQ. No.: 121-157; and (ii) a VH of (i) that contains one or more amino acid substitutions, deletions or insertions of a maximum of five, four, three, four, two or one amino acid; (b) one or more variable light chains (VLs) selected from the group consisting of: (i) ID. SEQ. No. 158-194, and (ii) a VL of (i) containing 25 one or more amino acid substitutions, deletions or insertions of a maximum of five, four, three, four, two or one amino acid; or (c) one or more variable heavy chains from (a) and one or more variable light chains from (b).

143

In another embodiment, the variable heavy chain (VH) has at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of sequence identity with a sequence of amino acids selected from the group consisting of the ID. SEQ. N °: 121-157 and / or the variable light chain (VL) has at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%. 98% or 99% sequence identity with an amino acid sequence selected from the group consisting of the ID. SEQ. No. 158-194.

In one aspect, an antigen binding protein is also provided that specifically binds to an epitope comprising one or more amino acid residues of FGFRlc, FGRF2C, FGFR3C and FGFR4.

In one aspect, an antigen-binding protein is also provided that specifically binds to an epitope comprising one or more β-Klotho amino acid residues.

In another aspect, an isolated antigen binding protein is also provided that specifically binds to an epitope comprising one or more amino acid residues of β-Klotho and one or more amino acid residues of FGFRlc, FGFR2c, FGFR3C or FGFR4.

In yet another embodiment, the isolated antigen-binding protein described hereinabove comprises a first amino acid sequence comprising at least one of the CDRH consensus sequences disclosed herein, and a second amino acid sequence comprising at least one of the consensus sequences of CDRL disclosed here.

144

In one aspect, the first amino acid sequence comprises at least two of the CDRH consensus sequences and / or the second amino acid sequence comprises at least two of the CDRL consensus sequences. In certain embodiments, the first and second sequences of amino acids are covalently linked to each other.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 146, the CDRH2 of the ID. SEQ. No. 133 and the CDRH1 of the ID. SEQ. No. 122, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 190, CDRL2 of the ID. SEQ. No. 176 and CDRL1 of the ID. SEQ. No.: 167.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 147, the CDRH2 of the ID. SEQ. No. 133 and the CDRH1 of the ID. SEQ. No. 122, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 189, the CDRL2 of the ID. SEQ. No. 176 and CDRL1 of the ID. SEQ. N °:

167.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 148, the CDRH2 of the ID. SEQ. No. 133 and the CDRH1 of the ID. SEQ. No. 122, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 188, the CDRL2 of the ID. SEQ. No. 176 and CDRL1 of the ID. SEQ. N °:

166.

145

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No.: 14 8, the CDRH2 of the ID. SEQ. No. 134 and the CDRH1 of the ID. SEQ. No. 122, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 188, the CDRL2 of the ID. SEQ. No. 176 and CDRL1 of the ID. SEQ. N °:

166.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 150, the CDRH2 of the ID. SEQ. No. 136 and the CDRH1 of the ID. SEQ. No. 124, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 183, CDRL2 of the ID. SEQ. No. 174 and the ID CDRL1. SEQ. N °:

161.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 152, the CDRH2 of the ID. SEQ. No. 138 and the CDRH1 of the ID. SEQ. No. 124, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 184, CDRL2 of the ID. SEQ. No. 173 and CDRL1 of the ID. SEQ. N °:

162.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 152, the CDRH2 of the ID. SEQ. No. 139 and the CDRH1 of the ID. SEQ. No. 124, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. N °: 186,

146 to CDRL2 of the ID. SEQ. No. 173 and CDRL1 of the ID. SEQ. N °:

162.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 151, the CDRH2 of the ID. SEQ. No. 137 and the CDRH1 of the ID. SEQ. No. 124, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No.: 186, the ID CDRL2. SEQ. No. 173 and CDRL1 of the ID. SEQ. No. 164.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 151, the CDRH2 of the ID. SEQ. No. 137 and the CDRH1 of the ID. SEQ. No. 124, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 182, the CDRL2 of the ID. SEQ. No. 173 and CDRL1 of the ID. SEQ. No. 160.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 153, the CDRH2 of the ID. SEQ. No. 140 and the CDRH1 of the ID. SEQ. No. 131, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 185, the CDRL2 of the ID. SEQ. No. 173 and CDRL1 of the ID. SEQ. N °:

163.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 153, the CDRH2 of the ID. SEQ. No. 140 and the CDRH1 of the ID. SEQ. N °: 125, and / or

The second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 185, the CDRL2 of the ID. SEQ. No. 173 and CDRL1 of the ID. SEQ. No. 163.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No.: 14 9, the CDRH2 of the ID. SEQ. No. 135 and the CDRH1 of the ID. SEQ. No.: 123, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No.: 181, CDRL2 of the ID. SEQ. No. 172 and the ID CDRL1. SEQ. No. 159.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 149, the CDRH2 of the ID. SEQ. No. 135 and the CDRH1 of the ID. SEQ. No.: 123, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No.: 187, CDRL2 of ID. SEQ. No. 175 and CDRL1 of the ID. SEQ. No. 165.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 145, the CDRH2 of the ID. SEQ. No. 132 and the CDRH1 of the ID. SEQ. No. 121, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 180, the ID CDRL2. SEQ. No. 171 and CDRL1 of the ID. SEQ. No. 158.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein

148 comprises the CDRH3 of the ID. SEQ. No. 154, the CDRH2 of the ID. SEQ. No. 133 and the CDRH1 of the ID. SEQ. No. 126, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No.: 191, CDRL2 of the ID. SEQ. No. 171 and CDRL1 of the ID. SEQ. N °:

168.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 157, the CDRH2 of the ID. SEQ. No. 144 and the CDRH1 of the ID. SEQ. No. 130, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 192, the CDRL2 of the ID. SEQ. No. 177 and CDRL1 of the ID. SEQ. N °:

169.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 155, the CDRH2 of the ID. SEQ. No. 135 and the CDRH1 of the ID. SEQ. No. 127, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No.: 193, CDRL2 of the ID. SEQ. No. 178 and the ID CDRL1. SEQ. N °:

170.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 156, the CDRH2 of the ID. SEQ. No. 142 and the CDRH1 of the ID. SEQ. No. 129, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 194, CDRL2 of the ID. SEQ. No. 173 and CDRL1 of the ID. SEQ. N °:

163.

149

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 156, the CDRH2 of the ID. SEQ. No. 141 and the CDRH1 of the ID. SEQ. No. 128, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 194, CDRL2 of the ID. SEQ. No. 173 and CDRL1 of the ID. SEQ. No. 163.

In an additional embodiment, the first amino acid sequence of the isolated antigen binding protein comprises the CDRH3 of the ID. SEQ. No. 156, the CDRH2 of the ID. SEQ. No. 143 and the CDRH1 of the ID. SEQ. No. 128, and / or the second amino acid sequence of the isolated antigen binding protein comprises CDRL3 of ID. SEQ. No. 194, CDRL2 of the ID. SEQ. No. 179 and CDRL1 of the ID. SEQ. No. 163.

In an additional embodiment, the antigen binding protein comprises at least two CDRH sequences from H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, Hll, H12, H13, H14 heavy chain sequences , H15, H16, H17 or H18, as shown in Table 4A. Again, in an additional embodiment, the antigen binding protein comprises at least two CDRL sequences of LI, L2, L3, L4, L5, L6, L7, L8, L9, LIO, Lil, L12, L13 , L14, L15, L16, L17 or L18, as shown in Table 4B. In an additional embodiment, the antigen-binding protein comprises at least two CDRH sequences of heavy chain sequences H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, Hll, H12, H13, H14, H15, H16, H17 or H18 as shown in Table 4Ά, and at least two CDRLs of

150 light chain sequences LI, L2, L3, L4, L5, L6, L7, L8, L9, IOL, Lil, L12, L13, L14, L15, L16, L17 or L18, as shown in Table 4B.

In yet another embodiment, the antigen-binding protein comprises the sequences of CDRH1, CDRH2 and CDRH3 of heavy chain sequences Hl, H2, H3, H4, H5, H6, H7, H8, H9, H10, Hll, H12, H13 , H14, H15, H16, H17 or H18 as shown in Table 4A. In yet another embodiment, the antigen binding protein comprises the CDRL1, CDRL2 and CDRL3 sequences of LI, L2, L3, L4, L5, L6, L7, L8, L9, LIO, Lil, L12, L13 , L14, L15, L16, L17 or L18, as shown in Table 4B.

In yet another embodiment, the antigen binding protein comprises all six CDRs of LI and Hl, or L2 and H2, or L3 and H3, or L3 and H4, or L4 and H5, or L5 and H6, or L6 and H7 , or L7 and H8, or L8 and H7, or L9 and H9, or L9 and H10, or LIO and Hll, or Lil and Hll, or L12 and H12, or L13 and H13, or L14 and H14, or L15 and H15 , or L16 and H16, or L17 and H17, or L18 and H18, as shown in Tables 4A and 4B.

Table 4A - Heavy chain sequences.

REFERENCE HEAVY CHAIN (H #) ID No. SEQ. TOTAL HEAVY HEAVY VARIABLE (VH #) ID No. SEQ. OF HEAVY VARIABLE ID No. SEQ. OF CDRH1 ID No. SEQ. OF CDRH2 ID No. SEQ. OF CDRH3 17C3 Hl 30 V _H 1 66 122 133 146 22H5 H2 31 V _h 2 67 122 133 147

151

16H7 H3 32 V _h 3 68 122 133 148 24H11 H4 33 V _h 4 69 122 134 148 18G1 H5 34 V _h 5 70 124 136 150 17D8 H6 35 V _h 6 71 124 138 152 26H11 H7 36 V _h 7 72 124 139 152 12E4 H8 37 V _h 8 73 124 137 151 12C11 H7 37 V _h 8 73 124 137 151 21H2 H9 38 V _h 9 74 131 140 153 21B4 H10 39 V _h 10 75 125 140 153 18B11.1 Hll 40 V _h 11 76 123 135 149 18B11.2 Hll 40 V _h 11 77 123 135 149 2 0D4 H12 41 V _h 12 78 121 132 145 46D11 H13 42 V _h 13 79 126 133 154 40D2 H14 46 V _h 14 80 130 144 157 39F7 H16 44 V _h 16 82 129 142 156 39F11 H17 43 V _h 17 83 128 141 156 37D3 H15 47 V _h 15 81 127 135 155 3 9G5 H18 45 V _h 18 84 128 143 156

Table 4B - light chain sequences.

REFERENCE HEAVY CHAIN (L #) ID No. SEQ. TOTAL LIGHT LIGHT VARIABLE (VH #) ID No. SEQ. LIGHT VARIABLE ID No. SEQ. DA CDRL1 ID No. SEQ. DA CDRL2 ID No. SEQ. FROM CDRL3 17C3 LI 48 V _L 1 85 167 176 190 22H5 L2 49 V _l 2 86 167 176 189

152

16H7 L3 50 V _l 3 87 166 176 188 24H11 L3 50 V _l 3 87 166 176 188 18G1 L4 51 V _l 4 88 161 174 183 17D8 L5 52 V _l 5 89 162 173 184 26H11 L6 53 V _l 6 90 162 173 186 12E4 L7 54 V _l 7 91 164 173 186 12C11 L8 55 V _l 8 92 160 173 182 21H2 L9 56 V _l 9 93 163 173 185 21B4 L9 56 V _l 9 93 163 173 185 18B11.1 IOL 57 V _L 10 94 159 172 181 18B11.2 Lll 58 V _L 11 95 165 175 187 2 0D4 L12 59 V _l 12 96 158 171 180 46D11 L13 60 V _l 13 97 168 171 191 4 0D2 L14 61 V _l 14 98 169 177 192 39F7 L16 63 V _l 16 100 163 173 194 37D3 L15 62 V _l 15 99 170 178 193 39F11 L17 64 V _l 17 101 163 173 194 3 9G5 L18 65 V _l 18 102 163 179 194

In one aspect, isolated antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 provided herein may be a monoclonal antibody, a polyclonal antibody, a recombinant antibody, a human antibody, a humanized antibody, a chimeric antibody, a multispecific antibody, or an antibody fragment thereof.

In another embodiment, the antibody fragment of the isolated antigen binding proteins provided herein may be a Fab fragment, a Fab 'fragment, an F (ab') ₂

153 fragment, an Fv fragment, a diabody, or a single chain antibody molecule.

In an additional embodiment, an isolated antigen-binding protein that specifically binds to (i) βKlotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 provided herein is a human antibody and can be of the type IgGl, IgG2, IgG3 or IgG4.

In another embodiment, an isolated antigen-binding protein that specifically binds to (i) βKlotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 comprises a light or heavy chain polypeptide as shown in Tables 1A-1B. In some embodiments, an antigen-binding protein that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one of FGFRlc, FGFR2C, FGFR3c and FGFR4 comprises a light variable or heavy variable domain, for example, those listed in Tables 2A-2B. In yet other embodiments, an antigen-binding protein that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one from FGFRlc, FGFR2c, FGFR3C and FGFR4 comprises one, two or three CDRHs or one, two or three CDRLs as shown in Tables 3A-3B, 4A-4B and Table 6C, infra . These antigen binding proteins and, in fact, any of the antigen binding proteins disclosed herein, can be PEGylated with one or more PEG molecules, for example, PEG molecules that have

154 is a molecular weight selected from the group consisting of 5 K, 10 K, 20 K, 40 K, 50 K, 60 K, 80 K, 100 K or more than 100 K.

In yet another aspect, any antigen-binding protein that specifically binds to (i) β-Klotho;

(ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 provided herein can be coupled to a labeling group and can compete for binding to the extracellular portion of (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 with an antigen binding protein from one of the isolated antigen binding proteins provided herein. In one embodiment, the isolated antigen-binding protein provided here can lower blood glucose levels, lower triglyceride and cholesterol levels, or improve other glycemic parameters and cardiovascular risk factors when administered to a patient.

As will be noted, for any antigen binding protein that comprises more than one CDR provided in Tables 3A-3B and 4A-4B, any combination of CDRs selected regardless of the disclosed sequences may be useful. In this way, antigen binding proteins can be generated with one, two, three, four, five or six of the independently selected CDRs. However, as will be noted by those skilled in the art, specific modalities generally use combinations of CDRs that are non-repetitive, for example,

155 antigen binding is generally not done with two CDRH2 regions, etc.

Some of the antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 which are provided herein will be discussed in more detail below.

Antigen binding proteins and binding epitopes and binding domains

When an antigen-binding protein is said to bind to an epitope on (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one of FGFRlc, FGFR2c, FGFR3C and FGFR4, or to the extracellular domain of β-Klotho, FGFRlc, FGFR2c, FGFR3c or FGFR4, for example, this means that the antigen binding protein specifically binds to a specified portion of (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. In some embodiments, for example, in certain cases where the antigen-binding protein binds only to FGFRlc or β-Klotho, the antigen-binding protein can specifically bind to a polypeptide consisting of specified residues (for example, a specified segment of β-Klotho, FGFRlc, FGFR2c, FGFR3c or FGFR4, such as those residues disclosed in Example 14). In other embodiments, for example, in certain cases where an antigen-binding protein interacts with both β-Klotho and with one or more of FGFRlc, FGFR2c, FGFR3c and FGFR4, the antigen-binding protein can bind to residues, strings of waste,

156 or regions in either β-Klotho or FGFRlc, FGFR2c, FGFR3c or FGFR4, depending on which receptor the antigen binding protein recognizes. In yet other embodiments, the antigen binding protein will bind to residues, sequences or residues or regions of a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4, for example, FGFRlc.

In any of the modalities presented above, an antigen-binding protein of this type does not need to come into contact with each residue of (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, or with the extracellular domain of the cited proteins or complexes; and not every single amino acid substitution or deletion within (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3C and FGFR4, or the extracellular domain of the cited proteins or complexes necessarily needs to significantly affect the binding affinity.

The epitope specificity and the binding domain (s) of an antigen binding protein can be determined by several methods. Some methods, for example, may use truncated portions of an antigen. Other methods use mutated antigen in one or more specific residues, for example, by using an alanine scan or arginine scan approach or by generating and studying chimeric proteins in which several domains, regions or amino acids are exchanged between two proteins (for example, forms of

157 mice and humans from one or more of the target antigens or proteins), or by protease protection assays.

Competition antigen binding proteins

In another aspect, antigen binding proteins are provided that compete with one of the antibodies or functional fragments exemplified by binding to (i) βKlotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. These antigen binding proteins can also bind to the same epitope as one of the antigen binding proteins exemplified herein, or to a superimposed epitope. Antigen binding proteins and fragments that compete or bind to the same epitope as the exemplified antigen binding proteins are expected to exhibit similar functional properties. The exemplary antigen-binding proteins and fragments include those with the H1-H18 and L1-L18 heavy and light chains, V _L 1V _l 18 and V _h 1-V _h 18 variable region domains and CDRs provided herein, including those in Tables 1, 2, 3 and 4. Thus, as a specific example, the antigen binding proteins that are provided include those that compete with an antibody that comprises:

(a) 1, 2, 3, 4, 5 or all 6 of the CDRs listed for an antibody listed in Tables 3A and 3B, and 4A and 4B, and Table 6C, infra;

(b) a V _H and a V _L selected from V _L 1-V _L 18 and V _H 1-V _H 18 and listed for an antibody listed in Tables 2A and 2B;

or

158 (c) two light chains and two heavy chains as specified for an antibody listed in Tables IA and 12B and Table 6A, infra.

Thus, in one embodiment, the present disclosure provides antigen binding proteins that compete for binding to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 with a reference antibody, wherein the reference antibody comprises a combination of sequences from the variable domain of the light chain and the heavy chain selected from the group consisting of L1H1, L2H2, L3H3, L3H4, L4H5, L5H6, L6H7, L7H8, L8H8, L9H9, L9H10, L10H11, L11H11, L12H12, L13H13, L14H14, L15H15, L17H16, L17 In another embodiment, the present disclosure provides human antibodies that compete for binding to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 with a reference antibody, wherein the reference antibody is 17C3, 22H5, 16H7, 24H11, 18G1, 17D8, 26H11, 12E4, 12C11, 21H2, 21B4, 18B11.1, 18B11.2, 20D4, 46D11, 40D2, 37D3, 39F7, 39F1 or 3 9G5.

In an additional embodiment, an isolated human antibody that binds to (i) β-Klotho is provided; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 with substantially the same Kd as a reference antibody; initiates FGF21-type signaling in an ELK-Luciferase assay in vitro to the same degree as a reference antibody; reduces blood glucose; reduces serum levels of

159 (j * lipids; and / or competes for binding with said J ”reference antibody to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4, where the reference antibody is selected from the group consisting of 17C3, 22H5, 16H7, 24H11, 18G1, 17D8, 26H11,

12E4, 12C11, 21H2, 21B4, 18B11.1, 18B11.2, 20D4, 46D11,

40D2, 37D3, 39F7, 39F1 or 39G5.

The ability to compete with an antibody can be determined using any suitable assay, for example, the one described in Example 8, in which 17C3, 22H5, 16H7, 24H11, 18G1, 17D8, 26H11, 12E4 antigen binding proteins, 12C11, 21H2, 21B4, 18B11.1, 18B11.2, 20D4,

46D11, 40D2, 37D3, 39F7, 39F1 or 39G5 can be used as the reference antibody.

Monoclonal antibodies

The antigen binding proteins that are provided include monoclonal antibodies that bind to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex 20 comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, and induce FGF21 type signaling to varying degrees. Monoclonal antibodies can be produced using any methodology known in the art, for example, by immortalizing splenic cells collected from the transgenic animal 25 after the immunization schedule has ended. Splenic cells can be immortalized using any methodology known in the art, for example, by fusing it with myeloma cells to produce hybridomas. Myeloma cells for use in fusion procedures 30 hybridoma producers are preferably non-producers

160 of antibody, have high fusion efficiency and enzyme deficiencies that make them unable to grow in certain selective media that support the growth of only the desired fused cells (hybridomas). Examples of cell lines suitable for use in mouse fusions include Sp-20, P3-X63 / Ag8, P3-X63-Ag8.653, NSl / l.Ag 4 1, Sp210-Agl4, FO, NSO / U, MPC -11, MPC11-X45-GTG 1.7 and S194 / 5XXO Bul; examples of cell lines used in mouse fusions include R210.RCY3, Y3-Ag 1.2.3, IR983F and 4B210. Other cell lines useful for cell fusions are U-266, GM1500-GRG2, LICR-LON-HMy2 and UC729-6.

In some cases, a hybridoma cell line is produced by immunizing an animal (for example, a transgenic animal that has human immunoglobulin sequences) with an FGFRlc, β-Klotho or FGFRlc and / or β-Klotho immunogen (by For example, a soluble complex comprising extracellular domains of FGFRlc, FGFR2c, FGFR3c or FGFR4 and / or β-Klotho, as shown in Examples 2 and 3; Membranes in which extracellular domains of FGFRlc, FGFR2c, FGFR3c or FGFR4 and / or β-Klotho are expressed, as shown in Examples 1 and 3, or whole cells that express FGFRlc and / or β-Klotho, as shown in Examples 1 and 3); collection of splenic cells from the immunized animal; fusion of the splenic cells collected to a myeloma cell line, thus generating hybridoma cells; establishment of hybridoma cell lines by hybridoma cells, and identification of a hybridoma cell line that produces an antibody that binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or

161 j FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 (for example, as described in Example 4) and can induce FGF21-type signaling (for example, as described in Examples 5-7) . These hybridoma cell lines, and the monoclonal antibodies produced by them, form aspects of the present disclosure.

Monoclonal antibodies secreted by a hybridoma cell line can be purified using any methodology known in the art. Hybridomas or mAbs can also be evaluated to identify mAbs with particular properties, for example, the ability to induce FGF21 type signaling. Examples of these assessments are provided here.

Chimeric and humanized antibodies

Chimeric and humanized antibodies based on the sequences presented above can easily be generated. One example is a chimeric antibody, which is an antibody composed of protein segments of different antibodies that are covalently joined to produce 20 functional immunoglobulin light or heavy chains or immunologically functional portions thereof. Generally, a

portion of jail heavy and / or light chain is identical or homologous The an sequence corresponding in antibodies derivatives in an species in particular or that belong to 25 a class or subclass antibody in particular,

while the rest of the chain (s) is identical or homologous to a corresponding sequence in antibodies derived from another species or belonging to another class or subclass of antibody. For methods related to chimeric antibodies, see, for example, U.S. Patent No. 4,816,567; and

162

Morrison et al., 198 5, Proc. Natl. Acad. Sci. USA 81: 6.851-6.855, which are incorporated herein by reference. The CDR graft is described, for example, in U.S. Patents No. 6,180,370, No. 5,693,762, No. 5,693,761, No. 5,585,089 and No. 5,530,101.

Generally, the goal of producing a chimeric antibody is to create a chimera in which the number of amino acids of the desired patient / recipient species is maximized. An example is the antibody with grafted CDR, where the antibody comprises one or more complementarity determining regions (CDRs) of a particular species or which belong to a particular antibody class or subclass, while the rest of the chain (s) (s) of the antibody is identical or homologous to a corresponding sequence in antibodies derived from another species or belonging to another class or subclass of antibody. For use in humans, the variable region or CDRs selected from a rodent antibody are often grafted onto a human antibody, replacing the naturally occurring variable regions or CDRs of the human antibody.

A useful type of chimeric antibody is a humanized antibody. Generally, a humanized antibody is produced by a monoclonal antibody first raised in a non-human animal. Certain amino acid residues in that monoclonal antibody, typically from antigen unrecognized portions of the antibody, are modified to be homologous to the corresponding residues in a corresponding isotype human antibody. Humanization can be accomplished, for

163 example, using various methods by replacing at least a portion of a variable rodent region with the corresponding regions of a human antibody (see, for example, US Patents No. 5,585,089 and No. 5,693,762; Jones et al., 1986, Nature 321: 522-525; Riechmann et al., 1988, Nature 332: 323-27; Verhoeyen et al., 1988, Science 239: 1.534-1.536).

In one aspect, the CDRs of the variable regions of the light and heavy chain of the antibodies provided here (for example, in Tables 3 and 4) are grafted into framework regions (FRs) of antibodies of the same phylogenetic species or of a different phylogenetic species. For example, the CDRs of the variable regions of the heavy and light chain V _H 15 V _H 2, V _H 3, V _h 4, V _h 5, V _h 6, V _h 7, V _h 8, V _h 9, V _H 10, V _H 11, V _H 12, V _H 13, V _H 14, V _h 15, V _h 16, V _h 17 or V _h 18 and / or V _L 1, V _L 2, V _L 3, V _L 4, V _L 5, V _L 6, V _l 7, V _l 8, V _l 9, V _l 10, V _l 11, V _h 12, V _l 13, V _l 14, V _L 15, V _L 16, V _l 17 or V _l 18 can be grafted into human consensual FRs. To create consensus human FRs, FRs from various amino acid sequences of the heavy or human light chain can be aligned to identify a consensus amino acid sequence. In other embodiments, the FRs of a heavy chain or light chain disclosed herein are replaced with the FRs of a different heavy chain or light chain. In one aspect, rare amino acids in the FRs of the heavy and light chains of an antigen-binding protein (for example, an antibody) that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one of FGFRlc, FGFR2c, FGFR3C and FGFR4 are not substituted, while the rest of the FR amino acids are

164 replaced. A rare amino acid is a specific amino acid that is in a position where that particular amino acid is not normally found in an RF. Alternatively, the grafted variable regions of a heavy or light chain can be used with a constant region that is different from the constant region of that particular heavy or light chain, as disclosed herein. In other embodiments, the variable regions grafted are part of a single chain Fv antibody.

In certain embodiments, constant regions of species other than human can be used together with the human variable region (or regions) to produce hybrid antibodies.

Fully human antibodies

Fully human antibodies are also provided by the present disclosure. Methods are available for producing fully human antibodies specific to a certain antigen, without exposing humans to the antigen (fully human antibodies). A specific means provided for implementing the production of fully human antibodies is the humanization of the humoral immune system of the mouse. The introduction of human immunoglobulin (Ig) loci in mice in which the endogenous Ig genes have been inactivated is a means of producing fully human monoclonal antibodies (mAbs) in mice, an animal that can be immunized with any desired antigen. The use of fully human antibodies can minimize the immunogenic and allergic responses that can sometimes be caused by

165 administration of mouse mAbs or mouse derivatives to humans as therapeutic agents.

Fully human antibodies can be produced by immunizing transgenic animals (typically mice) that are capable of producing a repertoire of human antibodies in the absence of endogenous immunoglobulin production. Antigens for this purpose typically have six or more contiguous amino acids and, optionally, are conjugated to a vehicle, for example, a hapten. See, for example, Jakobovits et al, (1993) Proc. Natl. Acad. Know. USA 90: 2.551-2.555; Jakobovits et al., (1993) Nature 362: 255-258; and Bruggermann et al., (1993) Year in Immunol. 7: 33. In an example of such a method, transgenic animals are produced by disabling the endogenous mouse immunoglobulin loci that encode the heavy and light chains of mouse immunoglobulin, and insertion into the mouse genome of large loci fragments that contain human genome DNA that encodes human heavy and light chain proteins. Partially modified animals, which have less than the total complement of human immunoglobulin loci, are then cross-mated to obtain an animal that has all the desired immune system modifications. When given the administration of an immunogen, these transgenic animals produce antibodies that are immunospecific for the immunogen, but have human, rather than murine, amino acid sequences, including variable regions. For more details on these methods, see, for example, WO 96/33735 and WO 94/02602. Additional methods related to

166 transgenic mice for the production of human antibodies are described in US Patents No. 5,545,807, No. 6,713,610, No. 6,673,986, No. 6,162,963, No. 5,545,807, No. 6,300,129, N 6,255,458, 5,877,397, 5,874,299 and 5,545,806; in PCT Publications WO 91/10741, WO 90/04036, and in EP 546073 BI and EP 546073 A1.

The transgenic mice described above, here called HuMab mice, contain a miniloccus of the human immunoglobulin gene that encodes unorganized heavy chain ([μ, mu] and [γ, gamma]) and light [K, kappa] immunoglobulin sequences human, along with targeted mutations that inactivate the endogenous loci of the μ [mu] and K [kappa] chain (Lonberg et al, 1994, Nature 368: 856-859). Consequently, mice exhibit reduced expression of mouse IgM or [k, kappa] in response to immunization, and the human heavy and light chain transgenes introduced undergo class change and somatic mutation to generate human IgG monoclonal antibodies [k, kappa] with high affinity (Lonberg et al., supra; Lonberg and Huszar, (1995) Intern. Rev. Immunol. 13: 65-93; Harding and Lonberg, (1995) Ann. NY Acad. Sci. 764: 536- 546). The preparation of HuMab mice is described in detail in Taylor et al., (1992) Nucleic Acids Research 20: 6.287-6.295; Chen et al. (1993) International Immunology 5: 647-656; Tuaillon et al., (1994) J. Immunol. 152: 2,912-2,920; Lonberg et al., (1994) Nature 368: 856859; Lonberg, (1994) Handbook of Exp. Pharmacology 113: 49101; Taylor et al., (1994) International Immunology 6: 579-591; Lonberg and Huszar, (1995) Intern. Rev. Immunol. 13: 65-93; Harding and Lonberg, (1995) Ann. N.Y. Acad. Sci. 764:

167

536-546; Fishwild et al., (1996) Nature Biotechnology 14: 845-851; the aforementioned references are hereby incorporated by reference in their entirety for all purposes. See also US Patents No. 5,545,806, No. 5,569,825, No. 5,625,126, No. 5,633,425, No. 5,789,650, No. 5,877,397, No. 5,661,016, No. 5,814 .318, No. 5,874,299, and No. 5,770,429; as well as US Patent No. 5,545,807; International Publication Number WO 93/1227; WO 92/22646; and WO 92/03918, the disclosures of which are hereby incorporated by reference in their entirety for all purposes. The technologies used for the production of human antibodies in these transgenic mice are also disclosed in WO 98/24893, and Mendez et al, (1997) Nature Genetics 15: 146-156, which are incorporated herein by reference. For example, the HCo7 and HCol2 transgenic mouse strains can be used to generate antigen-binding proteins (for example, antibodies) that bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4, and can induce FGF21-like signaling. More details regarding the production of human antibodies using transgenic mice are provided in the examples below.

Using hybridoma technology, antigen-specific human mAbs with the desired specificity can be produced and selected by transgenic mice, such as those described above. These antibodies can be cloned and expressed using a suitable vector and host cell, or the antibodies can be harvested from cultured hybridoma cells.

168

Fully human antibodies can also be derived from phage display libraries (as disclosed in Hoogenboom et al., (1991) J. Mol. Biol. 227: 381; and Marks et al., (1991) J. Mol. Biol. 222: 581). Phage presentation techniques mimic immune selection by presenting antibody repertoires on the surface of filamentous bacteriophages, and subsequent phage selection by binding to an antigen of choice. Such a technique is described in PCT Publication No. WO 99/10494 (incorporated herein by reference), which describes the isolation of high-affinity, functional agonist antibodies to MPL and msk receptors using such an approach.

Bispecific or bifunctional antigen binding proteins

Bispecific and bifunctional antibodies are also provided that include one or more CDRs or one or more variable regions, as described above. A bispecific or bifunctional antibody in some cases may be an artificial hybrid antibody that has two different heavy / light chain pairs and two different binding sites. Bispecific antibodies can be produced by a variety of methods, including, without limitation, fusion of hybridomas or ligation of Fab 'fragments. See, for example, Songsivilai and Lachmann, 1990, Clin. Exp. Immunol. 79: 315-321; Kostelny et al, 1992, J. Immunol. 148: 1.547-1.553. When an antigen binding protein of the present disclosure binds (i) both to β-Klotho and to one or more of FGFRlc, FGFR2C, FGFR3C or FGFR4; or (ii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c,

169

FGFR3c and FGFR4, the binding can lead to the activation of FGF21 type activity, as measured by the functional and signaling FGF21 type assays described in Examples 57; when such an antigen binding protein is an antibody, it is referred to as an agonist antibody.

Various other ways

Some of the antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 which are provided in the present disclosure include variant forms of the antigen binding proteins disclosed herein (for example, those having the sequences listed in Tables 1- 4).

In various embodiments, the antigen binding proteins disclosed herein can comprise one or more non-naturally occurring amino acids. For example, some of the antigen-binding proteins have one or more non-naturally occurring amino acid substitutions in one or more of the heavy or light chains, variable regions or CDRs listed in Tables 1-4. Examples of non-naturally occurring amino acids (which can replace any of the naturally occurring amino acids found in any sequence disclosed here, as desired) include: 4-hydroxyproline, γ-carboxyglutamate, ε-Ν, Ν, Νtrimethillisine, ε-Ν- acetillisin, O-phosphoserine, Nacetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylisine, σ-Ν-methylylginine, and other similar amino acids and imino acids (for example, 4-hydroxyproline). In the polypeptide annotation used here, the left direction

170 is the direction of the amino terminal and the right direction is the direction of the carboxyl terminal, according to standard usage and convention. A non-limiting list of examples of non-naturally occurring amino acids that can be inserted into an antigen-binding protein sequence or to replace a wild-type residue in an antigen-binding sequence includes β-amino acids, homoamino acids, cyclic amino acids and amino acids with derivatized side chains. Examples include (in L form or D form; abbreviated in parentheses): citrulline (Cit), homocitrulline (hCit), Να-methylcitrulin (NMeCit), Na-methylomocitrulin (Na-MeHoCit), ornithine (Orn), NaMethylornitine (Να -MeOrn or NMeOrn), sarcosine (Sar), homolysin (hLys or hK), homoarginine (hArg or hR), homoglutamine (hQ), Ncx-methylilarginine (NMeR), Namethilleucine (Na-MeL or NMeL), N-methylmethylamine ( NMeHoK), Να-methylglutamine (NMeQ), norleucine (Nle), norvaline (Nva), 1,2,3,4-tetrahydroisoquinoline (Tie), octahidroindol-2-carboxylic acid (Oic), 3- (1-naphthyl) alanine (1Nal), 3- (2-naphthyl) alanine (2-Nal), 1,2,3,4tetrahydroisoquinoline (Tie), 2-indanylglycine (Igl), paraiodophenylalanine (pl-Phe), para-aminophenylalanine (4AmP or 4-Amino-Phe), 4-guanidine phenylalanine (Guf), glycillisine (abbreviated K (Νε-glycyl) or K (glycyl) or K (gly)), nitrophenylalanine (nitrophe), aminophenylalanine (aminophe or Amino-Phe), benzylphenylalanine (benzylphe), γ-carboxyglutamic acid (γ-carboxylu) , hydroxyproline (hydroxypro), p-carboxyl-phenylalanine (Cpa), aaminoadypic acid (Aad), Να-methyl valine (NMeVal), N-a-methyl leucine (NMeLeu), Ncx-methylnorleucine (NMeNle),

171 cyclopentylglycine (Cpg), cyclohexylglycine (Chg), acetylarginine (acetilarg), a, β-diaminopropionic acid (Dpr), a, γ-diaminobutyric acid (Dab), diaminopropionic acid (Dap), cyclohexylalanine (Cha), 4-meth (MePhe), β, β-diphenyl-alanine (BiPhA), aminobutyric acid (Abu), 4-phenyl-phenylalanine (or biphenylalanine; 4Bip), α-amino-isobutyric acid (Aib), beta-alanine, beta-alanine aminopropionic, piperidinic acid, aminocaprioic acid, aminoheptanoic acid, aminopimelic acid, desmosine, diaminopimelic acid, Netylglycine, N-ethylparasparin, hydroxylsine, allohydroxylisin, isodesmosine, allo-isoleucine, methyl-glycol, N-methylamine; γ-carboxyglutamate, ε-Ν, Ν, Ν-trimethylsine, ε-Ν-acetillisine, O-phosphoserine, Nacetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylisin, G5-methyl-arginine, 4-amino-0A-phthalic acid and other similar amino acids, and derivatized forms of either those specifically listed.

In addition, antigen binding proteins can have one or more conservative amino acid substitutions in one or more of the heavy or light chains, variable regions or CDRs listed in Tables 1-4. Naturally occurring amino acids can be divided into classes based on common side chain properties:

1) hydrophobic: norleucine, Met, Ala, Val, Leu, Ile;

2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gin;

3) acids: Asp, Glu;

4) basic: His, Lys, Arg;

172

5) residues that influence the orientation of the chain: Gly, Pro; and

6) aromatics: Trp, Tyr, Phe.

Conservative amino acid substitutions may involve exchanging a member of one of these classes with another member of the same class. Conservative amino acid substitutions may include non-naturally occurring amino acid residues, which are typically incorporated by chemical peptide synthesis, rather than by synthesis in biological systems. See Table 5, below. These include peptidomimetics and other reverse or inverted forms of amino acid moieties. Non-conservative substitutions may involve exchanging a member of one of the above classes for a member of another class. Such substituted residues can be introduced into regions of the antibody that are homologous to human antibodies, or into the non-homologous regions of the molecule.

In producing these changes, according to certain modalities, the hydropathic index of amino acids can be considered. The hydropathic profile of a protein is calculated by assigning each amino acid a numerical value (hydropathy index) and then weighting those values repeatedly along the peptide chain. Each amino acid was assigned a hydropathic index based on its hydrophobicity and charge characteristics. These are: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine / cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (-0.7); serine (-0.8); tryptophan (-0.9); tyrosine (-1.3); proline (-1.6); histidine (-3.2); glutamate (-3.5);

173 glutamine (-3.5); aspartate (-3.5); asparagine (-3.5); lysine (-3.9); and arginine (-4.5).

The importance of the hydropathic profile in conferring interactive biological function on a protein is understood in the art (see, for example, Kyte et al., 1982, J. Mol. Biol. 157: 105-131). It is known that certain amino acids can replace other amino acids that have a similar hydropathic index or score and still retain a similar biological activity. In making changes based on the hydropathic index, in certain modalities, the substitution of amino acids whose hydropathic indexes are within ± 2 is included. In some ways, those that are within ± 1 are included, and in other ways, those that are within ± 0.5 are included.

It is also understood in the art that the substitution of similar amino acids can be done effectively on the basis of hydrophilia, particularly when the biologically functional protein or peptide thus created is intended for use in immunological modalities, as in the present case. In certain embodiments, the highest average local hydrophilicity of a protein, as determined by the hydrophilicity of its adjacent amino acids, is correlated with its immunogenicity and antigen or immunogenicity binding, that is, with a biological property of the protein.

The following hydrophilic values have been attributed to these amino acid residues: arginine (+3.0); lysine (+3.0); aspartate (+3.0 ± 1); glutamate (+3.0 ± 1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine (-0.4); proline (-0.5 ± 1); alanine (-0.5); histidine (-0.5); cysteine (-1.0); methionine (-1.3); valine

174 (-1.5); leucine (-1.8); isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5) and tryptophan (-3.4). In making changes based on similar hydrophilic values, in certain modalities, the substitution of amino acids whose hydrophilic values are within ± 2 is included, in other modalities, those which are within ± 1 are included, and in still others modalities, those that are within ± 0.5 are included. In some cases, it is also possible to identify epitopes of primary 10 amino acid sequences based on hydrophilia. These regions are also called central epitopic regions.

Conservative substitutions of exemplary amino acids are shown in Table 5.

Table 5 - Conservative amino acid substitutions.

Original waste Exemplary replacements Allah To be Arg Lys Asn Gin, His Asp Glu Cys To be Gin Asn Glu Asp Gly Pro His Asn, Gin Ile Read, Go Read Ile, go Lys Arg, Gin, Glu Met Leu, Ile Phe Met, Leu, Tyr To be Thr

175

Thr To be Trp Tyr Tyr Trp, Phe Val Ile, Leu

Those skilled in the art will be able to determine suitable polypeptide variants as presented herein using well known techniques associated with the information provided herein. Those skilled in the art can identify suitable areas of the molecule that can be altered without destroying the activity by targeting regions that are not supposed to be important to the activity. Those skilled in the art will also be able to identify residues and portions of the molecules that are conserved among similar polypeptides. In additional modalities, even areas that may be important for biological activity or structure may be subject to conservative amino acid substitutions, without destroying biological activity or without adversely affecting the structure of the polypeptide.

Additionally, those skilled in the art can review structure-function studies that identify residues in similar polypeptides that are important for the activity or structure. In the light of this comparison, one can predict the importance of amino acid residues in a protein that correspond to amino acid residues important for activity or structure in similar proteins. Those skilled in the art can opt for chemically similar amino acid substitutions for these presumably important amino acid residues.

176

Those skilled in the art can also analyze the three-dimensional structure and the sequence of amino acids in relation to that structure in similar polypeptides. In the light of this information, those skilled in the art can predict the alignment of amino acid residues in an antibody with respect to its three-dimensional structure. Those skilled in the art may choose not to make radical changes to amino acid residues that are presumably on the surface of the protein, insofar as these residues may be involved in important interactions with other molecules. In addition, those skilled in the art can generate test variants containing a single amino acid substitution in each desired amino acid residue. These variants can then be evaluated using tests for signaling type

FGF21 (see the Examples provided here), thus generating information regarding which amino acids can be changed and which should not be changed. In other words, based on the information gathered from these 20 routine experiments, those skilled in the art can easily determine the positions of amino acids where further substitutions must be avoided, either alone or in combination with other mutations.

Several scientific publications have dedicated themselves to predicting the secondary structure. See Moult, (1996) Curr.

Op. In Biotech. Ί_ · 422-427; Chou et al., (1974) Biochem. 13: 222-245; Chou et al., (1974) Biochemistry 113: 211222; Chou et al., (1978) Adv. Enzymol. Report Areas Mol. Biol. 47: 45-148; Chou et al., (1979) Ann. Rev. Biochem.

47: 251-276; and Chou et al., (1979) Biophys. J. 26: 367177

384. In addition, computer programs are currently available to assist with forecasting the secondary structure. One method of predicting the secondary structure is based on homology modeling. For example, two polypeptides or proteins that have a sequence identity greater than 30% or similarity greater than 40% can have similar structural topologies. The growth of the structural protein database (PDB) provided increased predictability of the secondary structure, including the potential number of folds within the structure of a polypeptide or protein. See Holm et al., (1999) Nucl. Acid. Res. 27: 244-247. It has been suggested (Brenner et al., (1997) Curr. Op. Struct. Biol.

Ί_ ·. 369-376) that there are a limited number of folds in a certain polypeptide or protein and that, once a critical number of structures have been resolved, the structural prediction will become dramatically more accurate.

Additional methods of forecasting the secondary structure include threading (Jones, (1997) Curr. Opin. Struct.

Biol. Ί_: 377-387; Sippl et al., (1996) Structure 4: 1519), profile analysis (Bowie et al., (1991) Science 253: 164-170; Gribskov et al., (1990) Meth. Enzym. 183: 146-159 ; Gribskov et al., (1987) Proc. Nat. Acad. Sci. 84: 4.355-4.358) and evolutionary link (see, Holm, (1999) 25 supra; and Brenner, (1997) supra).

In some modalities, amino acid substitutions are made that: (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) change the binding affinity to form protein complexes, (4) change affinities binder binding

178 or antigen and / or (4) confer or modify other physical-chemical or functional properties in these polypeptides. For example, single or multiple amino acid substitutions (in certain embodiments, conservative amino acid substitutions) can be made in the naturally occurring sequence. Substitutions can be made in that portion of the antibody that lies outside the domain (s) that form intermolecular contacts. In these modalities, conservative amino acid substitutions can be used that do not substantially alter the structural characteristics of the parent sequence (for example, one or more amino acid exchanges that do not disrupt the secondary structure that characterizes the parent or native antigen binding protein). Examples of secondary and tertiary polypeptide structures recognized in the art are described in Proteins, Structures and Molecular Principles (Creighton, Ed.), 1984, W.H. New York: Freeman and Company; Introduction to Protein Structure (Branden and Tooze, eds.), 1991, New York: Garland Publishing; and Thornton et al., (1991) Nature 3 54: 105, which are all incorporated herein by reference.

Additional preferred antibody variants include cysteine variants in which one or more cisterne residues in the parent or native amino acid sequence are deleted or replaced with another amino acid (e.g., serine). Cysteine variants are useful, inter alia, when antibodies must be refolded into a biologically active conformation. Cysteine variants may have less cysteine residues than the native antibody, and typically have an even number to minimize

179 interactions that result from unpaired cysteines.

The heavy and light chains, variable region domains and CDRs that are disclosed can be used to prepare polypeptides that contain an antigen binding region that can specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, and can induce FGF21-like signaling. For example, one or more of the CDRs listed in Tables 3 and 4 can be incorporated into a molecule (for example, a polypeptide) covalently or non-covalently to make an immunoadhesion. An immunoadhesion can incorporate a CDR (s) as part of a larger polypeptide chain, can covalently link one CDR (s) to another polypeptide chain, or can incorporate a CDR (s) non-covalently. The CDR (s) allows the immunoadhesion to bind specifically to a particular antigen of interest (for example, (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex which comprises βKlotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 or an epitope in them).

The heavy and light chains, variable region domains and CDRs that are disclosed can be used to prepare polypeptides that contain an antigen binding region that can specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, and can induce FGF21-like signaling. For example, one or more of the CDRs listed in Tables 3 and 4 can be incorporated into a molecule (for example, a

180 polypeptide) which is structurally similar to an antibody half comprising the heavy chain, the light chain of an antigen binding protein paired with an Fc fragment, such that the antigen binding region is monovalent (like a fragment) Fab), but with a dimeric Fc portion.

Mimetics (for example, peptide mimetics or peptidomimetics) are also provided based on the variable region domains and CDRs that are described herein. Such analogs can be peptides, non-peptides or combinations of peptide and non-peptide regions; Fauchere, 1986, Adv. Drug Res. 15:29; Veber and Freidinger, 1985, TINS page 392; and Evans et al, 1987, J. Med. Chem. 30: 1.22 9, which are incorporated herein by reference for any purpose. Peptide mimetics that are structurally similar to therapeutically useful peptides can be used to produce a similar therapeutic or prophylactic effect. These compounds are often developed with the aid of computerized molecular modeling. Generally, peptidomimetics are proteins that are structurally similar to an antibody that exhibits a desired biological activity, for example, the ability to specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, but have one or more peptide bonds optionally substituted by a selected link from: -CH ₂ NH-, CH ₂ S-, -CH2 -CH2-, -CH-CH- (cis and trans), -COCH ₂ -, -CH (OH) CH ₂ and -CH2SO-, by methods well known in the art. The systematic substitution of one or more amino acids in a

The consensus sequence with a D-amino acid of the same type (for example, D-lysine in place of L-lysine) can be used in certain modalities to generate more stable proteins. In addition, restricted peptides that comprise a consensus sequence or a variation of the substantially identical consensus sequence can be generated by methods known in the art (Rizo and Gierasch, 1992, Ann. Rev. Biochem. 61: 387, incorporated herein by reference) , for example, by adding internal cysteine residues capable of forming intramolecular disulfide bridges that cyclize the peptide.

Derivatives of antigen binding proteins that specifically bind to (i) βKlotho are also provided; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 which are described herein. Derivatized antigen binding proteins can comprise any molecule or substance that imparts a desired property to the antibody or fragment, for example, increased half-life in a particular use. The derivatized antigen-binding protein may comprise, for example, a detectable portion (or tag) (for example, a radioactive, colorimetric, antigenic or enzymatic molecule, a detectable globule (such as, for example, a magnetic or electrodense globule (eg gold)), or a molecule that binds to another molecule (for example, biotin or streptavidin), a therapeutic or diagnostic portion (for example, a radioactive, cytotoxic or pharmaceutically active portion), or a molecule that increases

182 suitability of the antigen binding protein for a particular use (for example, administration to an individual, for example, a human individual, or other uses in vivo or in vitro). Examples of molecules that can be used to derivatize an antigen-binding protein include albumin (for example, human serum albumin) and polyethylene glycol (PEG). Derivatives linked to albumin and PEGylates of antigen binding proteins can be prepared using methodologies well known in the art. Certain antigen binding proteins include a PEGylated single chain polypeptide, as described herein. In one embodiment, the antigen-binding protein is conjugated or otherwise linked to transthyretin (TTR) or a variant of TTR. The TTR or TTR variant can be modified chemically with, for example, a chemical substance selected from the group consisting of dextran, poly (n-vinyl pyrrolidone), polyethylene glycols, propropylene glycol homopolymers, polypropylene oxide / oxide copolymers ethylene, polyoxyethylated polyols and polyvinyl alcohols.

Other derivatives include covalent or aggregative conjugates of antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 which are disclosed herein with other proteins or polypeptides, for example, by expression of recombinant fusion proteins comprising heterologous polypeptides fused to the N or C terminus of an antigen-binding protein that induces FGF21-like signaling. For example,

183 the conjugated peptide may be a heterologous (or leader) signaling polypeptide, for example, the yeast alpha-factor leader, or a peptide such as, for example, an epitope tag. A fusion protein that contains an antigen-binding protein of the present disclosure can comprise peptides added to facilitate the purification or identification of an antigen-binding protein that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 (for example, a polyHis tag) and which can induce FGF21 type signaling. An antigen-binding protein that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 can also be linked to the FLAG peptide, as described in Hopp et al, 1988, Bio / Technology 6: 1,204; and U.S. Patent No. 5,011,912. The FLAG peptide is highly antigenic and provides an epitope reversibly linked to a specific monoclonal antibody (mAb), allowing rapid assay and easy purification of the expressed recombinant protein. Reagents useful for the preparation of fusion proteins in which the FLAG peptide is fused to a certain polypeptide are commercially available (Sigma, St. Louis, MO).

Multimers comprising one or more antigen-binding proteins that specifically bind to (i) βKlotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 form another aspect of the present disclosure. Multimers can take the form of dimers, trimers or

184 larger multimers attached covalently or non-covalently. Multimers comprising two or more antigen-binding proteins that bind to (i) βKlotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex that comprises β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 and that can induce FGF21 type signaling are contemplated for use as therapeutic, diagnostic substances and also for other uses, with an example of a multimer of this type being a homodimer. Other exemplary multimers include heterodimers, homotrimers, heterotrimers, homotetramers, heterotetramers, etc.

One modality is aimed at multimers that comprise multiple antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 joined via covalent or non-covalent interactions between peptide moieties fused to an antigen binding protein that specifically binds to (i) β -Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. These peptides can be peptide linkers (spacers), or peptides that have the property of promoting multimerization. Leucine zippers and certain antibody-derived polypeptides are among the peptides that can promote multimerization of antigen-binding proteins attached to them, as described in more detail here.

In particular modalities, multimers comprise

185 of two to four antigen binding proteins that bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. The antigen-binding protein portions of the multimer can be in any of the forms described above, for example, variants or fragments. Preferably, the multimers comprise antigen binding proteins that have the ability to specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4.

In one embodiment, an oligomer is prepared using immunoglobulin-derived polypeptides. The preparation of fusion proteins comprising certain heterologous polypeptides fused to various portions of antibody-derived polypeptides (including the Fc domain) has been described, for example, by Ashkenazi et al, (1991) Proc. Natl. Acad. Know. USA 88: 10.53 5; Byrn et al., (1990) Nature 344: 677; and Hollenbaugh et al, 1992 Construction of Immunoglobulin Fusion Proteins, in Current Protocols in Immunology, Supplement 4, pages 10.19.1-10.19.11.

One embodiment comprises a dimer comprising two fusion proteins created by fusing an antigen-binding protein that specifically binds to (i) βKlotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 to the Fc region of an antibody. The dimer can be made, for example, by inserting a gene fusion that encodes the fusion protein into an appropriate expression vector, which expresses the gene fusion in cells

186 hosts transformed with the recombinant expression vector, and which allows the expressed fusion protein to be assembled in a manner similar to antibody molecules, in which interchain disulfide bonds form between the Fc portions to generate the dimer.

The term Fc polypeptide, as used herein, includes native and mutein forms of polypeptides derived from the Fc region of an antibody. Truncated forms of these polypeptides that contain the hinge region that promotes dimerization are also included. Fusion proteins comprising Fc portions (and oligomers formed therefrom) offer the advantage of easy purification by affinity chromatography over Protein A or Protein G columns.

One suitable Fc polypeptide, described in PCT Application WO 93/10151 and US Patent ^Nos 5,426,048 and No. 5,262,522, is a single chain polypeptide extending from the terminal hinge region to the native C - C terminal Fc region of a human IgGl antibody. Another useful Fc polypeptide is the Fc mutein described in US Patent No. 5,457,035 and in Baum et al., (1994) EMBO J. 13: 3.992-4001. The amino acid sequence of this mutein is identical to that of the native Fc sequence presented in WO 93/10151, except that amino acid 19 has been changed from Leu to Ala, amino acid 20 has been changed from Leu to Glu and amino acid 22 has been changed from Gly to Ala. Mutein exhibits reduced affinity for Fc receptors.

In other embodiments, the variable portion of the heavy and / or light chains of an antigen binding protein such as that disclosed here may replace the portion

187 variable of an antibody heavy and / or light chain.

Alternatively, the oligomer is a fusion protein that comprises multiple antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, with or without peptide linkers (spacer peptides). Among the suitable peptide linkers are those described in US Patents ^Nos 4,751,180 and No. 4,935,233.

Another method for preparing oligomeric derivatives that comprise those antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 involves the use of a leucine zipper. Leucine zipper domains are peptides that promote the oligomerization of the proteins in which they are found. Leucine zippers were originally identified in several DNA-binding proteins (Landschulz et al., (1988) Science 240: 1759), and have since been found in several different proteins. Among the known leucine zippers are naturally occurring peptides and derivatives of these that dimerize or trimerize. Examples of leucine zipper domains suitable for the production of soluble oligomeric proteins are described in PCT Application WO 94/10308, and the leucine zipper derived from the lung D surfactant protein (SPD) surfactant described in Hoppe et al., (1994) FEBS Letters 344: 191, incorporated herein by reference. The use of a modified leucine zipper that allows stable trimerization of a heterologous protein to

188 he fused is described in Fanslow et al, (1994) Semin. Immunol. 6: 267-278. In one approach, recombinant fusion proteins that comprise an antigen-binding protein fragment or derivative that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 fused to a leucine zipper peptide are expressed in suitable host cells, and the fragments or derivatives of the soluble oligomeric antigen binding protein that they form are recovered from the culture supernatant.

In certain embodiments, the antigen binding protein has a K _D (equilibrium binding affinity) of less than 1 pM, 10 pM, 100 pM, 1 nM, 2 nM, 5 nM, 10 nM, 25 nM or 50 nM.

In another aspect, the present disclosure provides an antigen-binding protein that has a half-life of at least one day in vitro or in vivo (for example, when administered to a human subject). In one embodiment, the antigen-binding protein has a half-life of at least three days. In another embodiment, the antibody or portion thereof has a half-life of four days or more. In another embodiment, the antibody or portion thereof has a half-life of eight days or more. In another embodiment, the antibody or portion thereof has a half-life of ten days or more. In another embodiment, the antibody or portion thereof has a half-life of eleven days or more. In another embodiment, the antibody or portion thereof has a half-life of fifteen days or more. In another embodiment, the antibody or antigen binding portion thereof is

189 derivatized or modified in such a way that it has a longer half-life when compared to the non-derivatized or unmodified antibody. In another embodiment, an antigen-binding protein that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 contains point mutations to increase serum half-life, for example, as described in WO 00/09560, published on February 24, 2000 , incorporated by reference.

Glycosylation

An antigen-binding protein that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2C, FGFR3C and FGFR4 may have a glycosylation pattern that is different or altered from that found in the native species. As is known in the art, glycosylation patterns can depend both on the protein sequence (for example, on the presence or absence of particular glycosylation amino acid residues, discussed below) and on the host cell or organism in which the protein is produced. Particular expression systems will be discussed below.

Polypeptide glycosylation is typically N-linked or N-linked. N-linked refers to the adhesion of the carbohydrate moiety to the side chain of an asparagine residue. The asparagine-X-serine and asparagine-X-threonine tripeptide sequences, where X is any amino acid except proline, are the recognition sequences for enzymatic adhesion of the carbohydrate portion to the side chain

190 asparagine. Thus, the presence of one of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the adhesion of one of the N-acetylgalactosamine, galactose or xylose sugars to a hydroxyamino acid, more commonly serine or threonine, although 5-hydroxyproline or 5hydroxylysine can also be used.

The addition of glycosylation sites to the antigen-binding protein is conveniently achieved by altering 10 of the amino acid sequence so that it contains one or more of the tripeptide sequences described above (for N-linked glycosylation sites). The change can also be made by adding, or replacing, one or more serine or threonine residues to the starting sequence (for 15 glycosylation sites linked to O). To make it easier, the amino acid sequence of the antigen-binding protein can be altered through changes in the DNA level, particularly by mutating the DNA encoding the target polypeptide on such pre-selected bases

2 0 that are generated codons that will be translated we amino acids desired. Other means for increase the number of portions in carbohydrate in the protein binding antigen is per coupling chemical or enzymatic from glycosides The

protein. These procedures are advantageous, as they do not require the production of the protein in a host cell that has glycosylation capabilities for glycosylation linked to N and 0. Depending on the coupling mode used, the sugar (or sugars) can be attached 30 (a) arginine and histidine, (b) free carboxyl groups,

191 (c) free sulfhydryl groups, such as those of cysteine, (d) free hydroxyl groups, such as those of serine, threonine or hydroxyproline, (e) aromatic residues, such as those of phenylalanine, tyrosine or tryptophan, or (f) the amide group of glutamine. These methods are described in WO 87/05330 and in Aplin and Wriston, (1981) CRC Crit. Rev. Biochem. , pages 259-306.

The removal of carbohydrate moieties present in the starting antigen binding protein can be accomplished chemically or enzymatically. Chemical deglycosylation requires exposure of the protein to the compound trifluoromethanesulfonic acid, or an equivalent compound. This treatment results in the dividing of most or all sugars, except the binding sugar (Nacetylglucosamine or N-acetylgalactosamine), leaving the polypeptide intact. Chemical deglycosylation is described by Hakimuddin et al (1987) Arch. Biochem. Biophys. 259: 52 and by Edge et al., (1981) Anal. Biochem. 118: 131. Enzymatic dividing of carbohydrate moieties into polypeptides can be achieved by using several endo- and exo-glycosidases, as described by Thotakura et al., (1987) Meth. Enzymol. 138: 350. Glycosylation at potential glycosylation sites can be prevented by the use of the tunicamycin compound, as described by Duskin et al, (1982) J. Biol. Chem. 257: 3,105. Tunicamycin blocks the formation of protein-N-glycoside bonds.

Accordingly, aspects of the present disclosure include glycosylation variants of antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex that

192 comprises β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4, in which the number and / or type of glycosylation site (s) has been changed, compared with the amino acid sequences of the parent polypeptide. In certain embodiments, variants of the antibody protein comprise a greater or lesser number of N-linked glycosylation sites than the native antibody. An N-linked glycosylation site is characterized by the sequence: Asn-X-Ser or Asn-X-Thr, where the amino acid residue designated X can be any amino acid residue, except proline. Substituting amino acid residues to create this sequence provides a potential new site for the addition of an N-linked carbohydrate chain. Alternatively, substitutions that eliminate or alter this sequence will prevent the addition of an N-linked carbohydrate chain present in the polypeptide. native. For example, glycosylation can be reduced by deleting an Asn or replacing Asn with a different amino acid. In other modalities, one or more new sites linked to N are created. Antibodies typically have an N-linked glycosylation site in the Fc region.

Effector markers and groups

In some embodiments, an antigen-binding protein that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 comprises one or more markers. The term tag group or tag means any detectable tag. Examples of suitable labeling groups include, without limitation, the following: radioisotopes or

193 radionuclides (eg ³ H, ¹⁴ C, ¹⁵ N, ³⁵ S, ⁹⁰ Y, ⁹⁹ Tc, ^{1: L1} In, ¹²⁵ I, ¹³¹ I), fluorescent groups (eg, FITC, rhodamine, lanthanide matches), enzyme groups (eg, horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase), chemiluminescent groups, biotinyl groups or predetermined polypeptide epitopes recognized by a secondary reporter (for example, leucine zipper pair sequences, sites for secondary antibodies, meta-binding domains, epitope tags). In some embodiments, the labeling group is coupled to the antigen-binding protein by means of spacer arms of various lengths to reduce potential steric hindrance. Various methods for labeling proteins are known in the art and can be used, if considered suitable.

The term effector group means any group coupled to an antigen-binding protein that specifically binds to an (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 that acts as a cytotoxic agent. Examples for suitable effector groups are radioisotopes or radionuclides (for example, ³ H, ¹⁴ C, ¹⁵ N, ³⁵ S, ⁹⁰ Y, ⁹⁹ Tc, ^{1: L1} In, ¹²⁵ I, ¹³¹ I). Other suitable groups include toxins, therapeutic groups or chemotherapeutic groups. Examples of suitable groups include calicheamicin, auristatins, geldanamycin and cantansin. In some embodiments, the effector group is coupled to the antigen-binding protein by means of spacer arms of various lengths to reduce steric impediment

194 potential.

In general, markers are classified into several classes, depending on the assay in which they are to be detected: a) isotopic markers, which can be radioactive or heavy isotopes; b) magnetic markers (for example, magnetic particles); c) active redox portions; d) optical dyes; enzyme groups (for example, horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase); e) biotinylated groups; and f) predetermined polypeptide epitopes recognized by a secondary reporter (e.g., leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags, etc.). In some embodiments, the labeling group is coupled to the antigen-binding protein by means of spacer arms of various lengths to reduce potential steric hindrance. Various methods for labeling proteins are known in the art.

Specific markers include optical dyes, including, without limitation, chromophores, matches and fluorophores, with the latter being specific in many cases. Fluorophores can be small molecule fluorophores, or proteinaceous fluorophores.

The term fluorescent marker means any molecule that can be detected through its inherent fluorescent properties. Suitable fluorescent labels include, without limitation, fluorescein, rhodamine, tetramethylrodamine, eosin, erythrosine, coumarin, methyl coumarins, pyrene, Malachite green, stilbene, Lucifer Red, Cascade Blue,

195

Texas Red, IAEDANS, EDANS, BODIPY FL, LC Red 640, Cy 5, Cy 5.5, LC Red 705, Oregon Green, Alexa-Fluorine dyes (Alexa Fluorine 350, Alexa Fluorine 430, Alexa Fluorine 488, Alexa Fluorine 546, Alexa Fluorine 568, Alexa Fluorine 594, Alexa Fluorine 633, Alexa Fluorine 647, Alexa Fluorine 660, Alexa Fluorine 680), Cascade Blue, Cascade Yellow and R-phycoerythrin (PE) (Molecular Probes, Eugene, OR), FITC, Rhodamine and Red Texas (Pierce, Rockford, IL), Cy 5, Cy 5.5, Cy 7 (Amersham Life Science, Pittsburgh, PA). Suitable optical dyes, including fluorophores, are described in MOLECULAR PROBES HANDBOOK by Richard P. Haugland, hereby expressly incorporated by reference.

Suitable proteinaceous fluorescent markers also include, without limitation, green fluorescent protein, including a species of GFP Renilla, Ptilosarcus or Aequorea (Chalfie et al., (1994) Science 263: 802-805), EGFP (Clontech Labs., Inc. , Genbank Accession Number U55762), blue fluorescent protein (BFP, Quantum Biotechnologies, Inc., Quebec, Canada; Stauber, (1998) Biotechniques 24: 462-471; Heim et al., (1996) Curr. Biol. 6 : 178-182), enhanced yellow fluorescent protein (EYFP, Clontech Labs., Inc.), luciferase (Ichiki et al., (1993) J. Immunol. 150: 5.408-5.417), β-galactosidase (Nolan et al. , (1988) Proc. Natl. Acad. Sci. USA 85: 2,603-2,607) and Renilla (WO 92/15673, WO 95/07463, WO 98/14605, WO 98/26277, WO 99/49019, US Patents No. ° 5292658, N ° 5418155, N ° 5683888, N ° 5741668, N ° 5777079, N ° 5804387, N ° 5874304, N ° 5876995, N ° 5925558).

Preparation of antigen binding proteins

The non-human antibodies that are provided can

196 be, for example, derived from any antibody-producing animal, for example, mouse, rat, rabbit, goat, monkey, or non-human primate (such as, for example, monkey (e.g., Cynomolgus or Rhesus monkey) or simian ( for example, chimpanzees)). Non-human antibodies can be used, for example, in cell culture in vitro and in cell culture-based applications, or any other application in which an immune response to the antibody does not occur or is negligible, can be avoided, is not a concern, or is desired. In certain embodiments, antibodies can be produced by immunization with full-length βKlotho, FGFRlc, FGFR2c, FGFR3c or FGFR4 (Example 1), with the β-Klotho, FGFRlc, FGFR2C, FGFR3c or FGFR4 extracellular domain (Example 2), or two of βKlotho, FGFRlc, FGFR2c, FGFR3c and FGFR4 (Example 1), with whole cells that express FGFRlc, β-Klotho or both FGFRlc and β-Klotho (Examples 1 and 3), with membranes prepared from cells that express FGFRlc, βKlotho or both FGFRlc and β-Klotho (Example 1 and 3), with fusion proteins, for example, Fc fusions comprising FGFRlc, β-Klotho or FGFRlc and β-Klotho (or extracellular domains thereof) fused to Fc (Example 2 and 3), and other methods known in the art, for example, as described in the Examples presented herein. Alternatively, certain non-human antibodies can be raised by immunization with amino acids that are segments of one or more of β-Klotho, FGFRlc, FGFR2c, FGFR3c or FGFR4 that form part of the epitope to which certain antibodies provided herein bind. Antibodies can be polyclonal, monoclonal, or can be synthesized in

197 host cells by recombinant DNA expression.

Fully human antibodies can be prepared as described above by immunizing transgenic animals that contain human immunoglobulin loci or by selecting a phage display library that expresses a repertoire of human antibodies.

Monoclonal antibodies (mAbs) can be produced by a variety of techniques, including conventional monoclonal antibody methodology, for example, the standard somatic cell hybridization technique by Kohler and Milstein, (1975) Nature 256: 495. Alternatively, other techniques for the production of monoclonal antibodies can be employed, for example, viral or oncogenic transformation of B lymphocytes. A suitable animal system for the preparation of hybridomas is the murine system, which is a well-established procedure. Immunization protocols and techniques for the isolation of splenocytes immunized for fusion are known in the art. For these procedures, B cells from immunized mice are fused with a suitable immortalized fusion partner, for example, a murine myeloma cell line. If desired, rats or other mammals can be immunized instead of mice, and B cells from these animals can be fused with the murine myeloma cell line to form hybridomas. Alternatively, a myeloma cell line from a source other than mice can be used. Fusion procedures for the production of hybridomas are also well known. SLAM technology can also be used in the production of antibodies.

The single chain antibodies that are provided can

198 be formed by linking fragments of the variable domain of the heavy and light chain (Fv region) by means of an amino acid bridge (short peptide linker), resulting in a single polypeptide chain. These single chain Fvs (scFvs) can be prepared by fusing DNA that encodes a peptide linker between DNAs that encode the two polypeptides of the variable domain (V _L and V _H ). The resulting polypeptides can bend back over themselves to form antigen-binding monomers, or they can form multimers (for example, dimers, trimers, or tetramers), depending on the length of a flexible linker between the two variable domains (Kortt and cols., (1997) Prot. Eng. 10: 423; Kortt et al., (2001) Biomol. Eng. 18: 95-108). By combining different polypeptides that comprise V _L and V _H , multimeric scFvs can be formed that bind to different epitopes (Kriangkum et al., (2001) Biomol. Eng. 18: 31-40). Techniques developed for the production of single chain antibodies include those described in US Patent No. 4,946,778; Bird, (1988) Science 242: 423; Huston et al., (1988) Proc. Natl. Acad. Sci. USA 85: 5,879; Ward et al., (1989) Nature 334: 544, de Graaf et al., (2002) Methods Mol. Biol. 178: 379-387. Single chain antibodies derived from antibodies provided herein include, without limitation, scFvs that comprise variable domain combinations of the heavy and light chain variable regions disclosed in Table 2, or combinations of light and heavy chain variable domains that include CDRs disclosed in the Tables 3 and 4.

Antibodies provided here that are in a subclass can be changed to antibodies in a subclass

199 different using subclass change methods. In this way, IgG antibodies can be derived from an IgM antibody, for example, and vice versa. These techniques allow the preparation of new antibodies that have the antigen binding properties of a certain antibody (the antibody to the parent), but also exhibit biological properties associated with an isotype or subclass of antibody different from that of the parent antibody. Recombinant DNA techniques can be employed. The cloned DNA encoding particular polypeptides of the antibody can be employed in these procedures, for example, DNA encoding the constant domain of an antibody of the desired isotype. See, for example, Lantto et al., (2002) Methods Mol. Biol. 178: 303-316.

Accordingly, the antibodies that are provided include those that comprise, for example, the variable domain combinations described above, that have a desired isotype (for example, IgA, IgGl, IgG2, IgG3, IgG4, IgE and IgD), as well as Fab or F (ab ') ₂ fragments thereof. Moreover, if an IgG4 is desired, it may be desired to introduce a point mutation (CPSCP- 'CPPCP (IDS. SEQ. ^Nos 380-381, respectively, in order of occurrence)) in the hinge region as described in Bloom et al., (1997) Protein Science 6: 407, incorporated herein by reference) to alleviate the tendency to form intrachain H disulfide bonds that can lead to heterogeneity in IgG4 antibodies.

In addition, techniques for derivating antibodies that have different properties (ie, variable affinities for the antigen to which they bind) are also

200 known. One of these techniques, called chain shuffling, involves presenting the repertoire of the immunoglobulin variable domain gene on the surface of filamentous bacteriophage, often referred to as phage presentation. Chain shuffling has been used to prepare antibodies with high affinity to hapten 2-phenyloxazol-5-one, as described by Marks et al., (1992) BioTechnology 10: 779.

Conservative modifications were made to the variable regions of the heavy and light chain described in Table 2, or in the CDRs described in Tables 3A and 3B, 4A and 4B and Table 6C, infra (and corresponding modifications in the encoding nucleic acids) to produce a binding protein antigen that has functional and biochemical characteristics. Methods for obtaining these modifications are described above.

Proteins in Link in antigen what if call specifically to a or more in (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex what comprises β-Klotho and one of FGFRlc, FGFR2C, FGFR3C and FGFR4

they can also be modified in several ways. For example, if they are intended for therapeutic purposes, they can be conjugated to polyethylene glycol (PEGylated) to prolong the serum half-life or to increase the release of protein. Alternatively, the V region of the antibodies in question or fragments thereof can be fused to the Fc region of a different antibody molecule. The Fc region used for this purpose can be modified so that it does not bind to the complement, thereby reducing the likelihood of inducing cell lysis in the patient when

201 the fusion protein is used as a therapeutic agent. In addition, the antibodies in question or functional fragments thereof can be conjugated to human serum albumin to increase the serum half-life of the antibody or fragment thereof. Another useful fusion partner for antigen-binding proteins or fragments thereof is transthyretin (TTR). TTR has the ability to form a tetramer and, thus, an antibody fusion protein TTR can form a multivalent antibody that can increase its binding avidity.

Alternatively, substantial modifications in the functional and / or biochemical characteristics of the antigen binding proteins described herein can be obtained by creating substitutions in the amino acid sequence of the heavy and light chains that differ significantly in their effect on the maintenance of: (a) the structure the molecular framework in the substitution area, for example, as a lamina or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the strength of the side chain. A conservative amino acid substitution may involve a replacement of a native amino acid residue with a non-native residue that has little or no effect on the polarity or charge of the amino acid residue at that position. See Table 5, above. In addition, any native residue in the polypeptide can be replaced with alanine, as previously described for alanine scan mutagenesis.

Amino acid substitutions (whether conservative or non-conservative) of the antibodies in question can be implemented by those skilled in the art by

202 application of routine techniques. Amino acid substitutions can be used to identify important residues of the antibodies provided herein, or to increase or decrease the affinity of those antibodies for one or more of (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4, or for modifying the binding affinity of other antigen binding proteins described herein.

Methods of expressing antigen binding proteins

Expression systems and constructs in the form of plasmids, expression vectors, transcription or expression cassettes that comprise at least one polynucleotide as described above are also provided herein, as well as host cells that comprise such expression systems or constructs.

The antigen binding proteins provided herein can be prepared by a number of conventional techniques. For example, antigen binding proteins that specifically bind to (i) 13-10 Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 can be produced by recombinant expression systems, using any methodology known in the art. See, for example, Monoclonal Antibodies, Hybridomas: A New Dimension in Biological Analyzes, Kennet et al, (eds.) Plenum Press, New York (1980); and Antibodies: A Laboratory Manual, Harlow and Lane (eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1988).

203

Antigen binding proteins can be expressed in hybridoma cell lines (for example, in particular antibodies they can be expressed in hybridomas) or in cell lines other than hybridomas. Expression constructs that encode antibodies can be used to transform a mammalian, insect or microbial host cell. Transformation can be performed using any known method for introducing polynucleotides into a host cell including, for example, packaging the polynucleotide into a virus or bacteriophage and transducing a host cell with the construction by transfection procedures known in the art, as exemplified by US Patent No. 4,399,216; No. 4,912,040; No. 4,740,461; No. 4,959,455. The optimal transformation procedure used will depend on what type of host cell is being transformed. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art and include, without limitation, dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide (s) ) in liposomes, a mixture of nucleic acid with positively charged lipids and direct microinjection of DNA into the nuclei.

Recombinant expression constructs typically comprise a nucleic acid molecule that encodes a polypeptide that comprises one or more of the following: one or more CDRs provided herein; a light chain constant region; a variable region of the light chain; a heavy chain constant region (for example, C _H 1, C _H 2 and / or C _H 3);

204 and / or other portion of an antigen-binding protein framework. These nucleic acid sequences are inserted into an appropriate expression vector using standard ligation techniques. In one embodiment, the heavy or light chain constant region is attached to the C-terminus of the heavy or light chain variable region

anti-β-Klotho, -FGFRlc, -FGFR2C, -FGFR3C, -FGFR4 or β- Klotho and FGFRlc-specific and is turned on a vector in expression. 0 vector is typically selected for to be functional at host cell particular maid (or

that is, the vector is compatible with the machinery of the host cell, allowing the amplification and / or expression of the gene to occur). In some embodiments, vectors are used that employ protein-fragment complementation assays using protein reporters, for example, dihydrofolate reductase (see, for example, U.S. Patent No. 6,270,964, which is incorporated herein by reference). Suitable expression vectors can be purchased, for example, from Invitrogen Life Technologies or BD Biosciences (formerly Clontech). Other vectors useful for cloning and expressing antibodies and fragments include those described in Bianchi and McGrew, (2003) Biotech. Biotechnol. Bioeng. 84: 439-44, which is incorporated herein by reference. Additional suitable expression vectors are discussed, for example, in Methods Enzymol. , vol. 185 (D. V. Goeddel, ed.), 1990, New York: Academic Press.

Typically, the expression vectors used in any of the host cells will contain sequences for plasmid maintenance and for cloning and expression of exogenous nucleotide sequences. These strings,

205 collectively called flanking sequences, in certain embodiments will typically include one or more of the following nucleotide sequences: a promoter, one or more enhancer sequences, a source of replication, a transcription termination sequence, a complete intron sequence containing a site of donor and acceptor splice, a sequence encoding a leader sequence for polypeptide secretion, a ribosome binding site, a polyadenylation sequence, a multivinculator region for insertion of the nucleic acid encoding the polypeptide to be expressed, and a marker element selectable. Each of these sequences will be discussed below.

Optionally, the vector can contain a tag coding sequence, that is, an oligonucleotide molecule located at the 5 'or 3' end of an antigen binding protein coding sequence; the oligonucleotide sequence encodes polyHis (such as, for example, hexaHis (SEQ ID: NO: 382)), or another tag such as, for example, FLAG, HA (influenza virus hemagglutinin), or myc, for which antibodies are commercially available. This tag is typically fused to the polypeptide upon expression of the polypeptide, and can serve as a means for affinity purification or detection of the antigen binding protein by the host cell. Affinity purification can be achieved, for example, by column chromatography using antibodies against the tag as an affinity matrix. Optionally, the tag can subsequently be removed from the purified antigen binding protein by various means

206 such as, for example, the use of certain peptidases for cleavage.

Flanking sequences can be homologous (that is, of the same species and / or strain as the host cell), heterologous (that is, of a different species from the host cell species or strain), hybrids (that is, a combination of flanking sequences from more than one source), synthetic or native. Thus, the source of a flanking sequence can be any prokaryotic or eukaryotic organism, any vertebrate or invertebrate organism, or any plant, as long as the flanking sequence is functional, and can be activated, by the machinery of the host cell.

Flanking sequences useful in the vectors can be obtained by any of several methods well known in the art. Typically, flanking sequences useful in this application will have been previously identified by mapping and / or by restriction endonuclease digestion and can therefore be isolated from the appropriate tissue source with the use of appropriate restriction endonucleases. In some cases, the total nucleotide sequence of a flanking sequence may be known. Here, the flanking sequence can be synthesized using the methods described here for nucleic acid synthesis or cloning.

If all or only a portion of the flanking sequence is known, it can be obtained using polymerase chain reaction (PCR) and / or by screening a genomic library with a suitable probe, such as an oligonucleotide sequence fragment and / or flanker of the same or another species. When the

207 flanking sequence is not known, a DNA fragment containing a flanking sequence can be isolated from a larger piece of DNA that may contain, for example, a coding sequence or even another gene or genes. Isolation can be achieved by restriction endonuclease digestion to produce the appropriate DNA fragment, followed by isolation using agarose gel purification, Qiagen® column chromatography (Chatsworth, CA), or other methods known to those skilled in the art. The selection of suitable enzymes to achieve this objective will be easily evident to those skilled in the art.

An origin of replication is typically a part of those commercially acquired prokaryotic expression vectors, and the origin assists in amplifying the vector in a host cell. If the vector of choice does not contain a site of origin of replication, one can be synthesized chemically based on a known sequence, and ligated into the vector. For example, the origin of replication of plasmid pBR322 (New England Biolabs, Beverly, MA) is suitable for most gram-negative bacteria, and various viral origins (for example, SV40, polyoma, adenovirus, vesicular stomatitis virus (VSV ) or papillomaviruses such as HPV or BPV) are useful for cloning vectors in mammalian cells. Generally, the origin of replication component is not required for mammalian expression vectors (for example, the SV40 origin is often used only because it also contains the virus's early promoter).

A transcription termination sequence is

208 typically located 3 'to the end of a polypeptide coding region and serves to terminate transcription. Usually, a transcription termination sequence in prokaryotic cells is a fragment rich in G-C, followed by a poly-T sequence. Although the sequence is easily cloned from a library or even commercially acquired as part of a vector, it can also be easily synthesized using methods for nucleic acid synthesis, such as those described here.

A selectable marker gene encodes a protein necessary for the survival and growth of a host cell developed in a selective culture medium. Typical selection marker genes encode proteins that: (a) confer resistance to antibiotics or other toxins, for example, ampicillin, tetracycline or kanamycin for prokaryotic host cells; (b) complement the cell's auxotrophic deficiencies; or (c) supply crucial nutrients not available through complex or defined means. Specific selectable markers are the kanamycin resistance gene, the ampicillin resistance gene and the tetracycline resistance gene. Advantageously, a neomycin resistance gene can also be used for selection in both prokaryotic and eukaryotic host cells.

Other selectable genes can be used to amplify the gene that will be expressed. Amplification is the process whereby genes that are needed to produce a protein crucial to cell growth or survival are reiterated in tandem within the chromosomes of

209 successive generations of recombinant cells. Examples of selectable markers suitable for mammalian cells include dihydrofolate reductase (DHFR) genes and genes without a thymidine kinase promoter. Mammalian cell transformants are placed under selection pressure, in which only the transformants are exclusively adapted to survive due to the selectable gene present in the vector. The selection pressure is imposed by culturing the transformed cells under conditions in which the concentration of the selection agent in the medium is successively increased, thus leading to the amplification of both the selectable gene and the DNA encoding another gene, for example, a protein binding antigen that binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. As a result, increased amounts of a polypeptide, such as an antigen-binding protein, are synthesized by the amplified DNA.

A ribosome binding site is usually necessary for the initiation of mRNA translation and is characterized by a Shine-Dalgarno sequence (prokaryotes) or a Kozak sequence (eukaryotes). The element is typically located 3 'to the promoter and 5' to the coding sequence for the polypeptide to be expressed.

In some cases, for example, when glycosylation is desired in a eukaryotic host cell expression system, the various pre- or pro-consequences can be manipulated to increase glycosylation or yield. For example, you can change the dividing site of

210 peptidase of a particular signal peptide, or add pro-sequences, which can also affect glycosylation. The final protein product may have, at position -1 (relative to the first amino acid in the mature protein), one or more additional amino acids that affect expression, which may not have been completely removed. For example, the final protein product may have one or two amino acid residues found at the peptidase dividing site, attached to the amino terminus. Alternatively, the use of some enzyme dividing sites may result in a slightly truncated form of the desired polypeptide, if the enzyme cuts into such an area within the mature polypeptide.

Expression and cloning will typically contain a promoter that is recognized by the host organism and operationally linked to the molecule encoding an antigen binding protein that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. Promoters are non-transcribed sequences located upstream (that is, 5 ') of the start codon of a structural gene (usually within about 100 to 1,000 bp) that control the transcription of the structural gene. Promoters are conventionally grouped into one of two classes: inducible promoters and constitutive promoters. Inducible promoters initiate increased levels of DNA transcription under their control in response to some change in culture conditions, for example, the presence or absence of a nutrient or a change in temperature. Constitutive promoters, for

211 on the other hand, they uniformly transcribe a gene to which they are operationally linked, that is, with little or no control over gene expression. A large number of promoters, recognized by several potential host cells, are well known. A suitable promoter is operably linked to the DNA encoding the heavy chain or light chain comprising an antigen binding protein by removing the DNA promoter from the source by restriction enzyme digestion and inserting the desired promoter sequence into the vector.

Promoters suitable for use with yeast hosts are also well known in the art. Yeast enhancers are advantageously used with yeast promoters. Promoters suitable for use with mammalian host cells are well known and include, without limitation, those obtained from the genomes of viruses such as polyoma viruses, avian yaws viruses, adenovirus (such as, for example, Adenovirus 2), bovine papilloma, avian sarcoma virus, cytomegalovirus, retrovirus, hepatitis B virus and simian virus 40 (SV40). Other suitable mammalian promoters include heterologous mammalian promoters, for example, heat shock promoters and the actin promoter.

Additional promoters that may be of interest include, without limitation: SV40 early promoter (Benoist and Chambon, (1981) Nature 290: 304-310); CMV promoter (Thomsen et al., (1984) Proc. Natl. Acad. U.S.A. 81: 659663); the promoter contained in the 3 'long terminal repeat of the Rous sarcoma virus (Yamamoto et al., (1980) Cell 22: 787-797); herpes thymidine kinase promoter (Wagner and

212 cols., (1981) Proc. Natl. Acad. Sci. U.S.A. 78: 1.4441.445); promoter and regulatory sequences of the metallothionine gene (Prinster et al, (1982) Nature 296: 39-42); and prokaryotic promoters such as, for example, the beta-lactamase promoter (Villa-Kamaroff et al., (1978) Proc. Natl. Acad. Sci. U.S.A. 75: 3.727-3.731); or the tac promoter (DeBoer et al., (1983) Proc. Natl. Acad. Sci. U.S.A. 80: 21-25). Also of interest are the following animal transcription control regions, which exhibit tissue specificity and have been used in transgenic animals: the control region of the elastase I gene that is active in pancreatic acinar cells (Swift et al., (1984) Cell 38: 639-646; Omitz et al., (1986) Cold Spring Harbor Symp. Quant. Biol. 50: 399-409; MacDonald, (1987) Hepatology ·_ ·. 425-515); the control region of the insulin gene that is active in pancreatic beta cells (Hanahan, (1985) Nature 315: 115-122); the control region of the immunoglobulin gene that is active in lymphoid cells (Grosschedl et al., (1984) Cell 38: 647-658; Adames et al., (1985) Nature 318: 533-538; Alexander et al., (1987) Mol. Cell. Biol. Ί_ ·. 1,436-1,444); the control region of the mouse mammary tumor virus that is

active in cells testicular, breast, lymphoids and mast cells (Leder et al., (1986) Cell 45 : 485-495); The region of control of the gene of albumin that is active at the

liver (Pinkert et al., (1987) Genes and Devel. 1: 268276); the control region of the alpha-feto-protein gene that is active in the liver (Krumlauf et al., (1985) Mol.

Cell. Biol. 5: 1,639-1,648; Hammer et al., (1987) Science 253: 53-58); the alpha 1 gene control region

213 antitrypsin that is active in the liver (Kelsey et al, (1987) Genes and Devei. 1: 161-171); the control region of the beta-globin gene that is active in myeloid cells (Mogram et al., (1985) Nature 315: 338-340; Kollias et al., (1986) Cell 46: 89-94); the control region of the myelin basic protein gene that is active in oligodendrocyte cells in the brain (Readhead et al., (1987) Cell 48: 703-712); the control region of the myosin light chain-2 gene that is active in skeletal muscle (Sani, (1985) Nature 314: 283-286); and the gonadotropic release hormone gene control region that is active in the hypothalamus (Mason et al., (1986) Science 234: 1,372-1,378).

An enhancer sequence can be inserted into the vector to increase transcription of DNA encoding light or heavy chain that comprises an antigen binding protein that specifically binds to (i) βKlotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 by higher eukaryotes, for example, a human antigen binding protein that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. Intensifiers are cis-acting elements of DNA, usually about 10-300 bp in length, that act on the promoter to increase transcription. Intensifiers are relatively orientation and position-independent, having been found in both 5 'and 3' positions in relation to the transcription unit. Various intensifier sequences available

214 by mammalian genes are known (for example, globin, elastase, albumin, alpha-fetus-protein and insulin). Typically, however, a virus enhancer is used. The SV40 enhancer, cytomegalovirus early promoter enhancer, polyoma enhancer and adenovirus enhancers known in the art are exemplary enhancement elements for the activation of eukaryotic promoters. Although an enhancer can be positioned in the 5 'or 3' vector with respect to a coding sequence, it is typically located at a 5 'site of the promoter. A sequence that encodes an appropriate native or heterologous signal sequence (leader sequence or signal peptide) can be incorporated into an expression vector to promote extracellular secretion of the antibody. The choice of signal or leader peptide depends on the type of host cells in which the antibody is to be produced, and a heterologous signal sequence can replace the native signal sequence. Examples of signal peptides that are functional in mammalian host cells include the following: the signal sequence for interleukin-7 (IL-7) described in U.S. Patent No. 4,965,195; the signal sequence for the interleukin-2 receptor described in Cosman et al, (1984) Nature 312: 768; the interleukin-4 receptor signaling peptide described in EP Patent No. 0367 566; the interleukin-1 receptor type I signal peptide described in U.S. Patent No. 4,968,607; the interleukin-1 receptor type II signal peptide described in EP Patent No. 0 460 846.

The expression vectors that are provided can be

215 constructed from a starting vector, such as a commercially available vector. These vectors can contain, but not necessarily, all the desired flanking sequences. When one or more of the flanking sequences described here are no longer present in the vector, they can be obtained individually and linked in the vector. Methods used to obtain each of the flanking sequences are well known to those skilled in the art.

After the vector has been constructed and a nucleic acid molecule encoding a light chain, a heavy chain, or a light chain and a heavy chain comprising an antigen binding protein that specifically binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 having been inserted into the appropriate vector site, the completed vector can be inserted into a host cell suitable for amplification and / or expression of the polypeptide. Transformation of an expression vector into an antigen-binding protein in a selected host cell can be accomplished by well-known methods, including transfection, infection, calcium phosphate co-precipitation, electroporation, microinjection, lipofection, DEAE-mediated transfection -dextran, or other known techniques. The method selected will be, in part, a function of the type of host cell to be used. These methods and other suitable methods are well known to those skilled in the art, and are presented, for example, in Sambrook et al., (2001), supra.

216

A host cell, when grown under appropriate conditions, synthesizes an antigen-binding protein that can subsequently be harvested from the culture medium (if the host cell secretes it in the medium) or directly from the host cell that produces it (if not secreted) . The selection of an appropriate host cell will depend on several factors, for example, the levels of expression desired, the modifications of the polypeptide that are desirable or necessary for the activity (such as, for example, glycosylation or phosphorylation) and the ease of folding into one biologically active molecule.

Mammalian cell lines available as hosts for expression are well known in the art and include, without limitation, immortalized cell lines available from the American Type Culture Collection (ATCC), including, without limitation, Chinese hamster ovary (CHO) cells, HeLa cells, hamster cub kidney cells (BHK), monkey kidney cells (COS), human hepatocellular carcinoma cells (eg, HepG2), and several other cell lines. In certain embodiments, cell lines can be selected by determining which cell lines have high levels of expression and constitutively produce antigen binding proteins with desirable binding properties (for example, the ability to bind to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3C OR FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4). In another embodiment, a cell line of the B cell line that does not make its own antibody, but has an ability to make and secrete

217 a heterologous antibody, can be selected. The ability to induce FGF21-type signaling can also form a selection criterion.

Uses of antigen binding proteins for diagnostic and therapeutic purposes

The antigen binding proteins disclosed herein are useful for detecting (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 in biological samples and identification of cells or tissues that produce one or more of (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. For example, the antigen binding proteins disclosed herein can be used in diagnostic assays, for example, binding assays to detect and / or quantify (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 expressed in a tissue or cell. Antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 can be used to treat diseases related to FGF21 type signaling in a patient who needs it, for example, type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease and metabolic syndrome. By forming a signaling complex that comprises an antigen binding protein, and (i) βKlotho; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c,

218, -¼

FGFR3c and FGFR4, the natural FGF21 activity in vivo, which is associated with FGFRlc, FGFR2c, FGFR3C, FGFR4 and β-Klotho in vivo to initiate signaling, can be mimicked and / or increased, leading to therapeutic effects.

Indications

A disease or condition associated with human FGF21 includes any disease or condition whose appearance in a patient is caused, at least in part, by the induction of FGF21 type signaling, which is initiated in vivo by the formation of a complex comprising FGFRlc, FGFR2c, FGFR3c or FGFR4 and β-Klotho and FGF21. The severity of the disease or condition can also be reduced by inducing FGF21-like signaling. Examples of diseases and conditions that can be treated with antigen-binding proteins include type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease and metabolic syndrome.

The antigen-binding proteins described herein can be used to treat type 2 diabetes, obesity, dyslipidemia, NASH, cardiovascular disease and metabolic syndrome, or can be used as a prophylactic treatment administered, for example, daily, weekly, every two weeks. , monthly, bimonthly, biannually etc. to avoid or reduce the frequency and / or severity of symptoms, for example, high blood glucose levels, high levels of triglycerides and cholesterol, thereby providing an improved glycemic and cardiovascular risk factor profile.

Diagnostic methods

The antigen binding proteins described herein can be used for diagnostic purposes to detect,

219 diagnose or monitor diseases and / or conditions associated with FGFRlc, FGFR2c, FGFR3c, FGFR4, β-Klotho, FGF21 or combinations thereof. Methods are also provided for detecting the presence of (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 in a sample using classical immunohistological methods known to those skilled in the art (eg, Tijssen, 1993, Practice and Theory of Enzyme Immunoassays, Vol 15 (Eds RH Burdon and PH van Knippenberg, Elsevier, Amsterdam); Zola, (1987) Monoclonal Antibodies: A Manual of Techniques, pages 147-158 (CRC Press, Inc.); Jalkanen et al., (1985) J Cell, Biol. 101: 976-985; Jalkanen et al., (1987) J. Cell, Biol. 105: 3.087-3.096). The detection of (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and a FGFRlc, FGFR2C, FGFR3C and FGFR4 urn can be performed in vivo or in vitro.

The diagnostic applications provided here include the use of antigen binding proteins to detect the expression of (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 and / or the bond to (i) βKlotho; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4. Examples of methods useful in detecting the presence of (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 include immunoassays, for example, the enzyme linked immunosorbent assay (ELISA) and

220 the radioimmunoassay (RIA).

For diagnostic applications, the antigen binding protein will typically be tagged with a detectable tagging group. Suitable labeling groups include, without limitation, the following: radioisotopes or radionuclides (e.g., ³ H, ¹⁴ C, ¹⁵ N, ³⁵ S, ⁹⁰ Y, ⁹⁹ Tc, ^ In, ¹²⁵ I, ¹³¹ I) fluorescent groups ( e.g., FITC, rhodamine, phosphorus lanthanides), enzyme groups (e.g., horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase), chemiluminescent groups, predetermined biotinyl groups or polypeptide epitopes recognized by a secondary reporter ( for example, leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, 15 epitope tags). In some embodiments, the labeling group is coupled to the antigen-binding protein by means of spacer arms of various lengths to reduce potential steric hindrance. Various methods for labeling proteins are known in the art and 20 can be used.

In another aspect, an antigen-binding protein can be used to identify a cell or cells that express (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of 25 FGFRlc, FGFR2c, FGFR3c and FGFR4. In a specific embodiment, the antigen binding protein is labeled with a labeling group and the binding of the labeled antigen binding protein to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β30 Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 is detected.

221

In an additional specific embodiment, the binding of the antigen-binding protein to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 is detected in vivo. In an additional specific embodiment, the antigen binding protein is isolated and measured using methodologies known in the art. See, for example, Harlow and Lane, (1988) Antibodies: A Laboratory Manual, New York: Cold Spring Harbor (ed. 1991 and periodic supplements); John E. Coligan, ed. , (1993) Current Protocols In Immunology, New York: John Wiley & Sons.

Another aspect allows the detection of the presence of a test molecule that competes for binding to (i) β-Klotho;

(ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 with the provided antigen binding proteins, as disclosed herein. An example of such an assay would involve detecting the amount of free antigen binding protein in a solution that contains an amount of one or more of (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 in the presence or absence of the test molecule. An increase in the amount of free antigen binding protein (i.e., antigen binding protein not bound to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4) would indicate that the test molecule is capable of competing for binding to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3C or

222

FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 with the antigen binding protein. In one embodiment, the antigen-binding protein is tagged with a tagging group. Alternatively, the test molecule is labeled and the amount of free test molecule is monitored in the presence and absence of an antigen-binding protein.

Treatment methods: pharmaceutical formulations and routes of administration

Methods of using antigen binding proteins are also provided. In some methods, an antigen-binding protein is provided to a patient. The antigen binding protein induces FGF21-like signaling.

Pharmaceutical compositions are also provided which comprise a therapeutically effective amount of one or more of the antigen binding proteins and a pharmaceutically acceptable diluent, carrier, solubilizer, emulsifier, preservative and / or adjuvant. In addition, methods of treating a patient by administering this pharmaceutical composition are included. The term patient includes human patients.

Suitable formulation materials are non-toxic to recipients at the dosages and concentrations used. In specific embodiments, pharmaceutical compositions are provided that comprise a therapeutically effective amount of human antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4.

223

In certain embodiments, acceptable formulation materials are preferably non-toxic to the recipients at the dosages and concentrations employed. In certain embodiments, the pharmaceutical composition may contain formulation materials for modification, maintenance or conservation, for example, pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, dissolution or release rate, adsorption or penetration composition. In such embodiments, suitable formulation materials include, without limitation, amino acids (such as, for example, glycine, glutamine, asparagine, arginine or lysine); antimicrobials; antioxidants (such as, for example, ascorbic acid, sodium sulfite or sodium hydrogen sulfite); buffers (such as, for example, borate, bicarbonate, Tris-HCl, citrates, phosphates or other organic acids); bulking agents (such as, for example, mannitol or glycine); chelating agents (such as, for example, ethylenediamine tetraacetic acid (EDTA)); complexing agents (such as, for example, caffeine, polyvinylpyrrolidone, beta-cyclodextrin or hydroxypropyl beta-cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as, for example, glucose, mannose or dextrins); proteins (such as, for example, serum albumin, gelatin or immunoglobulins); coloring, flavoring and thinning agents; emulsifying agents; hydrophilic polymers (such as, for example, polyvinylpyrrolidone); low molecular weight polypeptides; salt-forming counter ions (such as sodium); preservatives (such as benzalkonium chloride, benzoic acid, salicylic acid,

224 thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid or hydrogen peroxide); solvents (such as, for example, glycerin, propylene glycol or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as, for example, Pluronics, PEG, sorbitan esters, polysorbates such as, for example, polysorbate 20, polysorbate, Triton, tromethamine, lecithin, cholesterol, tiloxapal); stability-enhancing agents (such as, for example, sucrose or sorbitol); tonicity-enhancing agents (such as, for example, alkali metal halides, preferably sodium or potassium chloride, mannitol sorbitol); release vehicles; diluents; pharmaceutical excipients and / or adjuvants. See Remington's Pharmaceutical Sciences, 18th Edition, (AR Gennaro, ed.), 1990, Mack Publishing Company.

In certain embodiments, the optimal pharmaceutical composition will be determined by those skilled in the art, depending, for example, on the desired route of administration, the release format and the desired dosage. See, for example, Remington's Pharmaceutical Sciences, supra. In certain embodiments, these compositions can influence the physical state, stability, in vivo release rate and in vivo clearance rate of the disclosed antigen binding proteins. In certain embodiments, the primary vehicle or carrier in a pharmaceutical composition can be aqueous or non-aqueous in nature. For example, a suitable vehicle or carrier can be water for injection, saline solution or cerebrospinal fluid

225 artificial, possibly supplemented with other common materials in compositions for parenteral administration. Neutral buffered saline or saline mixed with serum albumin are additional exemplary vehicles. In specific embodiments, the pharmaceutical compositions comprise Tris buffer with a pH of about 7.0-8.5, or acetate buffer with a pH of about 4.0-5.5, and can further include sorbitol or a suitable substitute. In certain embodiments, compositions that comprise antigen binding proteins that specifically bind to (i) βKlotho; (ii) FGFRlc, FGFR2c, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 can be prepared for storage by mixing the selected composition that has the desired degree of purity with optional formulation agents (Remington's Pharmaceutical Sciences, supra) in the form of a lyophilized cake or an aqueous solution. In addition, in certain embodiments, the antigen-binding protein that binds to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 can be formulated as a lyophilisate using appropriate excipients such as, for example, sucrose.

Pharmaceutical compositions can be selected for parenteral release. Alternatively, the compositions can be selected for inhalation or for delivery through the digestive tract, for example, orally. The preparation of such pharmaceutically acceptable compositions is within the skill of the art.

The formulation components are present

226 preferably in concentrations that are acceptable at the site of administration. In certain embodiments, buffers are used to maintain the composition at a physiological pH or at a slightly lower pH, typically within a pH range of about 5 to about 8.

When parenteral administration is contemplated, therapeutic compositions can be provided in the form of a parenterally acceptable aqueous solution, free of pyrogen, comprising the desired antigen binding protein in a pharmaceutically acceptable carrier. A particularly suitable vehicle for parenteral injection is sterile distilled water in which the antigen-binding protein is formulated as a sterile, properly preserved isotonic solution. In certain embodiments, the preparation may involve formulating the desired molecule with an agent, for example, injectable microspheres, bioerosible particles, polymeric compounds (such as, for example, polylactic acid or polyglycolic acid), globules or liposomes, which can provide controlled release or sustained product that can be released through deposit injection. In certain modalities, hyaluronic acid can also be used, which can have the effect of promoting sustained duration in the circulation. In certain embodiments, the implantable delivery of pharmacological devices can be used to introduce the desired antigen-binding protein.

Certain pharmaceutical compositions are formulated for inhalation. In some embodiments, antigen-binding proteins that bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex that comprises β

227

Klotho and one from FGFRlc, FGFR2c, FGFR3C and FGFR4 are formulated as an inhalable dry powder. In specific embodiments, solutions for inhaling the antigen-binding protein can also be formulated with a propellant for aerosol delivery. In certain embodiments, solutions can be nebulized. Pulmonary administration and formulation methods are therefore further described in International Patent Application No. PCT / US94 / 001875, which is incorporated by reference and describes the pulmonary release of chemically modified proteins. Some formulations can be administered orally. Antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2C, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 that are administered in this way can be formulated with or without vehicles normally used in the composition of solid dosage forms such as tablets and capsules . In certain embodiments, a capsule can be designed to release the active portion of the formulation at the point in the gastrointestinal tract when bioavailability has been maximized and pre-systemic degradation is minimized. Additional agents can be included to facilitate the absorption of an antigen-binding protein. Thinners, flavorings, low melting waxes, vegetable oils, lubricants, suspending agents, tablet disintegrating agents and binders can also be used.

Some pharmaceutical compositions comprise an effective amount of one or more binding proteins

228 human antigens that specifically bind to (i) βKlotho; (ii) FGFRlc, FGFR2C, FGFR3C or FGFR4; or (iii) a complex comprising β-Klotho and one of FGFRlc, FGFR2c, FGFR3c and FGFR4 in a mixture with non-toxic excipients that are suitable for the manufacture of tablets. By dissolving the tablets in sterile water, or in another appropriate vehicle, solutions in unit dose form can be prepared. Suitable excipients include, without limitation, inert diluents, for example, calcium carbonate, sodium carbonate or bicarbonate, lactose or calcium phosphate; or binding agents, for example, starch, gelatin, or acacia; or lubricating agents such as, for example, magnesium stearate, stearic acid or talc.

Additional pharmaceutical compositions will be evident to those skilled in the art, including formulations that involve antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising β-Klotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 in sustained or controlled release formulations. Techniques for formulating various other means of sustained or controlled release, for example, liposome vehicles, bioerosible microparticles or porous globules and deposit injections, are also known to those skilled in the art. See, for example, International Patent Application No. PCT / US93 / 00829, which is incorporated by reference and describes the controlled release of porous polymeric microparticles for the release of pharmaceutical compositions. Sustained-release preparations may include polymeric matrices

229 semipermeable in the form of molded articles, for example, films or microcapsules. Sustained-release matrices may include polyesters, hydrogels, polylactides (as disclosed in US Patent No. 3,773,919 and European Patent Application Publication No. EP 058481, each incorporated herein by reference), L-glutamic acid copolymers and gamma ethyl-L-glutamate (Sidman et al., 1983, Biopolymers 2: 547-556), poly (2-hydroxyethylmethacrylate) (Langer et al., 1981, J. Biomed. Mater. Res. 15: 167-277 and Langer, 1982, Chem. Tech. 12: 98-105), ethylene vinyl acetate (Langer et al, 1981, supra) or poly-D (-) - 3-hydroxybutyric acid (European Patent Application Publication No. EP 133,988). Sustained-release compositions can also include liposomes that can be prepared by any of several methods known in the art. See, for example, Eppstein et al, 1985, Proc. Natl. Acad. Sci. USA 82: 3,688-3,692; European Patent Application Publication ^Nos EP 036 676; EP 088,046 and EP 143,949, incorporated herein by reference.

Pharmaceutical compositions used for in vivo administration are typically supplied as sterile preparations. Sterilization can be achieved by filtration through sterile filtration membranes. When the composition is freeze-dried, sterilization using this method can be carried out before or after freeze-drying and reconstitution. Compositions for parenteral administration can be stored in lyophilized form or in a solution. Parenteral compositions are usually placed in a container that has a sterile access port, for example, a pouch or bottle of solution

230 IV which has a cap that can be pierced by a hypodermic injection needle.

In certain embodiments, cells that express a recombinant antigen-binding protein as disclosed herein are encapsulated for release (see, Invest. Ophthalmol. Vis. Sci. (2002) 43: 3.292-3.298 and Proc. Natl. Acad. Sciences USA ( 2006) 103: 3.896-3.901).

In certain formulations, an antigen-binding protein has a concentration of at least 10 mg / ml, 20 mg / ml, 30 mg / ml, 40 mg / ml, 50 mg / ml, 60 mg / ml, 70 mg / ml , 80 mg / ml, 90 mg / ml, 100 mg / ml or 150 mg / ml. Some formulations contain a buffer, sucrose and polysorbate. An example of a formulation is one containing 50-100 mg / ml of antigen binding protein, 5-20 mM sodium acetate, 5-10% w / v sucrose and 0.002-0.008% w / v polysorbate . Certain formulations, for example, contain 65-75 mg / ml of an antigen binding protein in 9-11 mM sodium acetate buffer, 8-10% w / v sucrose and 0.0050.006% w / v of polysorbate. The pH of some of these formulations is in the range of 4.5-6. Other formulations have a pH of 5.0-5.5 (for example, pH of 5.0, 5.2 or 5.4).

After the pharmaceutical composition has been formulated, it can be stored in sterile vials as a solution, suspension, gel, emulsion, solid, crystal or with a dehydrated or lyophilized powder. These formulations can be stored in a ready-to-use form or in a form (for example, lyophilized) that is reconstituted before administration. Kits for the production of a single dose administration unit are also provided. Certain

231 kits contain a first container that has a dry protein and a second container that has an aqueous formulation. In certain embodiments, kits are provided containing syringes pre-filled with a single or multiple chambers (for example, liquid syringes and syringes). The therapeutically effective amount of a pharmaceutical composition that contains antigen binding protein to be employed will depend, for example, on the context and therapeutic objectives. Those skilled in the art will note that the appropriate dosage levels for treatment will vary, depending in part on the molecule released, the indication for which the antigen binding protein is being used, the route of administration and the size (body weight) , body surface or organ size) and / or condition (age and general health) of the patient. In certain modalities, the doctor may titrate the dosage and modify the route of administration to obtain the optimal therapeutic effect.

A typical dosage can range from about 1 pg / kg to

about 30 mg / kg or more depending on factors mentioned above. In modalities specific, The dosage may vary from 10 pg / kg up until about 30 mg / kg, optionally from 0.1 mg / kg up until about 30 mg / kg, alternatively from 0, 3 mg / kg up until about 20 mg / kg. In

For some applications, the dosage is 0.5 mg / kg to 20 mg / kg. In some cases, an antigen-binding protein is dosed at 0.3 mg / kg, 0.5 mg / kg, 1 mg / kg, 3 mg / kg, 10 mg / kg or 20 mg / kg. The dosage dosage in some treatment regimens is at a dose of 0.3 mg / kg per week, 0.5 mg / kg per week, 1 mg / kg per week, 3 mg / kg per week,

232 mg / kg per week or 20 mg / kg per week.

The dosing frequency will depend on the pharmacokinetic parameters of the antigen binding protein in particular in the formulation used. Typically, a physician will administer the composition until a dosage is reached that achieves the desired effect. The composition can, therefore, be administered as a single dose, or as two or more doses (which may not necessarily contain the same amount of the desired molecule) over time, or as a continuous infusion through a delivery device. implantation or catheter. Appropriate dosages can be verified using appropriate dosage response data. In certain embodiments, antigen-binding proteins can be administered to patients over an extended period of time. Chronic administration of an antigen binding protein minimizes the adverse immune or allergic response commonly associated with antigen binding proteins that are not entirely human, for example, an antibody raised against a human antigen in a non-human animal, for example, a non-human antibody or non-human antibody produced in a non-human species.

The route of administration of the pharmaceutical composition is according to known methods, for example, orally, by injection through the intravenous, intraperitoneal, intracerebral (intraparenchymal), intracerebroventricular, intramuscular, intraocular, intraarterial, intraportal or intralesional route; sustained release systems or implantation devices. In certain embodiments, the compositions can be

233 administered by bolus injection or continuously by infusion, or by implantation device.

The composition can also be administered locally by implanting a membrane, sponge or other appropriate material on which the desired molecule has been absorbed or encapsulated. In certain embodiments, when an implantation device is used, the device can be implanted in any suitable tissue or organ, and the release of the desired molecule can occur through diffusion, controlled time bolus or continuous administration.

It may also be desirable to use pharmaceutical antigen-binding protein compositions ex vivo. In such cases, cells, tissues or organs that have been removed from the patient are exposed to the antigen-binding protein pharmaceutical compositions and then the cells, tissues and / or organs are subsequently implanted back into the patient.

In particular, antigen binding proteins that specifically bind to (i) β-Klotho; (ii) FGFRlc, FGFR2c, FGFR3c or FGFR4; or (iii) a complex comprising βKlotho and one from FGFRlc, FGFR2c, FGFR3c and FGFR4 can be released by implanting certain cells that have been genetically modified, using methods such as those described herein, to express and secrete the polypeptide. In certain embodiments, these cells may be animal or human cells, and may be autologous, heterologous, or xenogenic. In certain embodiments, cells may be immortalized. In other embodiments, in order to decrease the likelihood of an immune response, cells may be immortalized.

234 be encapsulated to prevent infiltration of surrounding tissues. In additional embodiments, encapsulation materials are typically biocompatible, semi-permeable polymeric attachments or membranes that allow the release of the protein product (s), but prevent the destruction of cells by the patient's immune system or other harmful factors in the surrounding tissues.

Combined therapies

In another aspect, the present disclosure provides a method of treating an individual for diabetes with a therapeutic antigen binding protein of the present disclosure, for example, the fully human therapeutic antibodies described herein, together with one or more other treatments. In one embodiment, such a combined therapy has an additive or synergistic effect. Antigen-binding proteins can be administered in combination with one or more of the currently available treatments for type 2 diabetes or obesity. These treatments for diabetes include biguanide (metaformin) and sulfonylureas (such as glyburide, glipizide). Additional treatments aimed at maintaining glucose homeostasis include PPAR gamma agonists (pioglitazone, rosiglitazone); glinides (meglitinide, repaglinide and nateglinide); DPP-4 inhibitors (Januvia® and Onglyza®) and alpha-glycosidase inhibitors (acarbose, voglibose).

Additional combined treatments for diabetes include injectable treatments such as insulin and incretin mimetics (Byetta®, Exenatide®), other GLP-1 (glucagon-like peptide) analogs such as

235 example, liraglutide, other GLP-1R and Symlin® agonists (pranlintide).

Additional combined treatments aimed at weight loss include Meridia® and Xenical®.

EXAMPLES

The following examples, including the experiments carried out and the results obtained, are provided for illustrative purposes only, and should not be considered as limiting.

EXAMPLE 1

PREPARATION OF CELLS THAT OVEREXPRESS FGFRlc FOR USE AS AN ANTIGEN

Nucleic acid sequences encoding the full-length human FGFRlc polypeptide (SEQ ID NO: 4; Figures 1A-1B) and a separate sequence encoding the full-length human β-Klotho polypeptide (ID. DE SEQ NO: 7; Figures 2A-2C) have been subcloned into suitable mammalian cell expression vectors (for example, pcDNA3.1 Zeo, pcDNA3.1 Hyg (Invitrogen, Carlsbad, CA) or pDSR «20. pDSR «20 contains an SV40 early promoter / enhancer for expression of the gene of interest and a mouse DHFR expression cassette for selection in CHO DHFR (-) host cells, such as AM1 CHO (a derivative of DG44, CHO DHFR (-)) ·

AM-1 CHO cells were seeded at 1.5 x 10 ⁶ cells per 100 mm plate. After 24 hours, the cells were cotransfected with linearized DNAs from pDSRa20 / huFGFRlc and pDSR «20 / h ^ -Klotho with FuGeneô (Roche Applied Science). The transfected cells were trypsinized 2 days after

236 transfection and sown in CHO DHFR selective growth medium containing 10% dialysed FBS and without hypoxanthine / thymidine supplement. After 2 weeks, the resulting transfected colonies were trypsinized and pooled.

HEK293T cells were transfected with huFGFRlc and full-length hup-Klotho in vector based on the pcDNA3.1 or pTT14 series (an expression vector developed by Durocher, NRCC, with CMV promoter and EBV ori, similar to pTT5 and an puromycin selection marker) and selected with the corresponding drugs after a procedure similar to that for CHO transfection and selection.

The pools of transfected AM1 CHO or 293T cells with FGF21R (i.e. FGFRlc and pKlotho) were repeatedly classified using Alexa 647-labeled FGF21. As a cell surface staining reagent, FGF21 was tagged with Alexa 647-NHS according to the method recommended by the manufacturer (Molecular Probes, Inc. Cat A 2006). Alexa 647-labeled FGF21 showed specific staining of cells expressing the FGF21R receptor and not of untransfected parental cells (Figure 3). Highly expressed cells were collected at the end of the final classification, expanded and frozen in flasks. AM-1 / huFGF21R cells were prepared for immunization and 293T / huFGF21R cells were used for titration of mouse sera by FACS after immunization and in binding assessments of hybridoma supernatants by FMAT (see Example 4).

EXAMPLE 2

237

PREPARATION OF A SOLUBLE FGFRlc / β-KLOTHO COMPLEX FOR USE AS AN ANTIGEN

Soluble FGF21 receptor constructs were generated in expression vectors pTT14 or pcDNA3.1. The FGFRlc ECD-Fc construct (SEQ ID NO: 362, Figure 4) comprises the N-terminal extracellular domain of FGFRlc (amino acid residues # 1-374; SEQ ID NO: 5) fused to Fc (SEQ ID. N °: 384). The β-Klotho ECD-Fc construct (SEQ ID. NO: 363, Figure 5) comprises the N-terminal extracellular domain of β-Klotho (amino acid residues # 1-996; SEQ ID. N °: 8) fused to Fc (SEQ ID. NO .: 384).

HEK293 cells (293F, Invitrogen) were transfected with ECD-Fc / pTT5 from huFGFRlc, ECD-Fc / pTT14-pure from ΙιηβKlotho and dGFP / pcDNA3.1-Neo and selected in the presence of the corresponding drugs, followed by repeated FACS separation based on dGFP expression. The cells were developed in Dulbecco's modified Eagle's medium (DMEM) free of serum supplemented with non-essential amino acids in HyperFlasks (Corning) for 4 days and conditioned media (CM), and collected for purification.

The 293 CM was concentrated 6 times and applied to Protein A FF balanced in PBS. The protein was eluted with Ag / Ab Pierce Gentle elution buffer. The Protein A pool was dialyzed against 20 mM Tris-HCl, pH 7, 10 mM NaCl and applied to SP HP at pH 7.0. The FGFRlc ECD-Fc was present in the flow (FT) and the heterodimer was eluted with a linear gradient of 0-0.4 M NaCl, 20 mM Tris-HCl pH 7.0. N-terminal amino acid sequencing found that the purified soluble FGF21R is a heterodimer

238 composed of a ratio (1: 1) of ECF-Fc of FGFRlc and ECDFc of β-Klotho. The purified soluble FGF21R-FC (Figure 6) was used as the antigen for immunization.

EXAMPLE 3

PREPARATION OF MONOCLONAL ANTIBODIES

Immunizations were performed using one or more suitable forms of the FGF21 receptor antigen, including: (1) cell-bound receptor for full-length human CHF transfectors and β-Klotho on the cell surface obtained by transfection of CHO cells with cDNA encoding a full-length human FGFRlc polypeptide from ID. SEQ. No. 4 (see also Figures la-b) and cDNA encoding a human β-Klotho polypeptide from ID. SEQ. N °: 7 (see also Figures 2a-c); (2) membrane extract from the aforementioned cells that express the FGF21R receptor complex; or (3) soluble FGF21R receptor that can be obtained by coexpression of the FGFRlc N-terminal extracellular domain (ECD) (SEQ ID NO: 5; see also Figure 4) and the N-terminal extracellular domain (ECD) ) of β-Klotho (SEQ ID. NO: 8; see also Figure 5) or (4) combinations of these.

An adequate amount of immunogen (i.e. 10 pg / mouse soluble FGF21R or 3-4 x 10 ⁶ cells / mouse of stably transfected CHO cells or 150 pg / mouse of purified FGF21R membranes prepared by CHO cells that stably express FGF21R) was used for initial immunization in XenoMouse ™ according to the methods disclosed in US Patent Application Serial No. 08 / 759,620, filed on 3

239 December 1996 and International Patent Application ^Nos WO 98/24893 and WO 00/76310, the disclosures of which are incorporated herein by reference. After the initial immunization, subsequent immunogen booster immunizations (5 pg / mouse soluble FGF21R or 1.7 x 10 ^s cells transfected with FGF21R / mouse or 75 pg of purified FGF21R membranes) were administered in a dosage and for the required duration to induce adequate antiFGF21R titration in mice. Titrations were determined by a suitable method, for example, by enzyme immunoassay, fluorescence activated cell separation (FACS), or by other methods (including combinations of enzyme immunoassays and FACS).

Animals that exhibit adequate titrations were identified, and lymphocytes were obtained from drainage lymph nodes and, if necessary, pooled for each cohort. Lymphocytes were dissociated from lymphoid tissue by trituration in a suitable medium (for example, Dulbecco's modified Eagle's medium; DMEM; obtained by Invitrogen, Carlsbad, CA) to release cells from the tissues, and suspended in DMEM. B cells were selected and / or expanded using standard methods, and fused to a suitable fusion partner, for example, myeloma non-secreting cells P3X63Ag8.653 (American Type Culture Collection CRL 1580; Kearney et al, J. Immunol. 123, 1979, 1.5481.550), using techniques that were known in the art.

In a suitable fusion method, lymphocytes were mixed with fusion partner cells in a 1: 4 ratio. The cell mixture was gently pelleted by centrifugation at 400 x g for 4 minutes, the supernatant

240 decanted, and the cell mixture gently mixed (for example, using a 1 ml pipette). The fusion was induced with PEG / DMSO (polyethylene glycol / dimethyl sulfoxide; obtained from Sigma-Aldrich, St. Louis MO; 1 ml per million lymphocytes). PEG / DMSO was added slowly with gentle stirring over one minute, followed by one minute of mixing. IDMEM (DMEM without glutamine; 2 ml per million B cells) was then added over 2 minutes with gentle agitation, followed by additional IDMEM (8 ml per million B cells) which was added over 3 minutes.

The fused cells were pelleted (400 xg for 6 minutes) and resuspended in 20 ml of selection medium (for example, DMEM containing Azaserine and Hypoxanthine [HA] and other supplementary materials, as needed) per million B cells. The cells were incubated for 20-30 minutes at 37 ° C and then resuspended in 200 ml of selection medium and cultured for three to four days in T175 flasks before plating in 96 wells.

The cells were distributed in 96-well plates using standard techniques to maximize the clonality of the resulting colonies. After several days of culture, supernatants were collected and subjected to evaluation assays, as detailed in the examples below, including confirmation of binding to the human FGF21 receptor, specificity and / or cross-reactivity between species. Positive cells were also selected and subjected to standard cloning and subcloning techniques. The clonal strains were expanded in vitro, and secreted human antibodies were obtained for analysis.

241

Thus, mice were immunized with cells or membranes that express full-length FGF21R cells, or soluble FGF21R extracellular domain, with a range of 11-17 immunizations over a period of approximately one month to three and a half months. Several cell lines that secrete specific antibodies to FGF21R have been obtained, and the antibodies have been further characterized. Their sequences are presented here and in the Sequence List, and the results of various tests using these antibodies are provided.

EXAMPLE 4

SELECTION OF FMAT BINDING ANTIBODIES

After 14 days of culture, hybridoma supernatants were evaluated for specific monoclonal antibodies to FGF21R by fluorimetric microvolumetric assay technology (FMAT) by evaluation against the CHO AMl / huFGF21R cell line or recombinant HEK293 cells that were transfected with human FGF21R and counter-evaluation against parental CHO or HEK293 cells. Briefly, cells in Freestyle medium (Invitrogen) were seeded in 384-well FMAT plates at a volume of 50 μΐ / well at a density of 4,000 cells / well for stable transfectants, and at a density of 16,000 cells / well for the parental cells, and the cells were incubated overnight at 37 ° C. Twenty-five μΐ / well of supernatant was then added, and the plates were incubated for approximately one hour at 4 ° C, and then 10 μΐ / well of secondary anti-human IgG-Cy5 antibody was added at a concentration of 2.8

242 pg / ml (400 ng / ml final concentration). The plates were then incubated for one hour at 4 ° C, and the fluorescence was read using a FMAT System cell detection system (Applied Biosystems).

In total, more than 3,000 hybridoma supernatants have been identified as binding to cells expressing the FGF21 receptor, but not to parental cells by the FMAT method. These supernatants were then tested in FGF21 functional assays, as described below.

EXAMPLE 5

SELECTION OF ANTIBODIES INDUCING FGF21 TYPE SIGNALING

Experiments were carried out to identify antibodies

functional what mimic activity in FGF21 like wild (per example, the skill for induce signaling of type FGF21) using a reporter rehearsal of

Suitable FGF21. The revealed FGF21 reporter assay measures the activation of FGFR signaling by reading the MAPK pathway. β-Klotho is a co-receptor for FGF21 signaling and, although it is believed to have no inherent signaling capacity due to its very short cytoplasmic domain, it is necessary for FGF21 to induce signaling through FGFRs.

Example 5.1

ELK-luciferase reporter-essay

ELK-luciferase assays were performed using a 293T renal cell system or recombinant human CHO cells. Specifically, the host cells were genetically modified to overexpress β-Klotho and luciferase reporter constructs. At

243 reporter constructs contain strings encoding GAL4-ELK1 and 5xUAS-Luc, a luciferase reporter conducted by a promoter that contains five tandem copies of the Gal4 binding site. Activation of the FGF21 receptor complex in these reporter strains of recombinant cells induces intracellular signal transduction which, in turn, leads to ERK and ELK phosphorylation. Luciferase activity is regulated by the level of phosphorylated ELK, and is used to indirectly monitor and quantify FGF21 activity.

In one example, CHO cells were transfected sequentially using the Lipofectamine 2000 transfection reagent (Invitrogen) according to the manufacturer's protocol with the receptor constructs expressing β-Klotho, FGFRlc and the reporter plasmids: 5xGal4Luciferase (minimal TK promoter with upstream 5xGal4 binding sites of luciferase) and Gal4-ELK1. Gal4-ELK1 binds to Gal4 binding sites and activates transcription when it is phosphorylated by ERK. Luciferase transcription and, therefore, the corresponding enzyme activity in this context, is regulated by the level of phosphorylated ELK1, and is used to indirectly monitor and quantify FGF21 activity.

Clone 2E10 was selected as the FGF21 luciferase reporter cell line based on the optimal 10-20 fold assay window, with native FGF21 exhibiting an EC ₅ o in the range of a single nM.

For the assay, ELK-luciferase reporter cells were plated on 96-well assay plates, and deprived of serum overnight. FGF21 or test samples were added for 6 hours at 37 ° C. It was allowed,

244 then, the plates were allowed to cool to room temperature and the luciferase activity in the cell lysates was measured with Bright-Glo (Promega).

Example 5.2

ERK phosphorylation test

An alternative host cell line, specifically L6 (a mouse myoblast cell line) was developed and applied to identify antibodies with FGF21-like signaling activity. The rat L6 cell line is a desirable host cell line for the activity assay, as it is known to express minimum levels of endogenous FGF receptors. L6 cells do not respond to FGF21, even when transfected with β-Klotho expression vector and, therefore, provide a cleaner background level (Kurosu et al., (2007) J. Biol. Chem. 282, 26,687- 26,695).

L6 cells were maintained in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum and penicillin / streptomycin. The cells were transfected with plasmids that express β-Klotho and individual FGFR using the transfection reagent Lipofectamine 2000 (Invitrogen) according to the manufacturer's protocol.

The analysis of FGF signaling in L6 cells was performed as described in the literature (Kurosu et al., (2007) J. Biol. Chem. 282, 26,687-26,695). Cell cultures were collected 10 min after the treatment of FGF21 or test molecules and suddenly frozen in liquid nitrogen, homogenized in the lysis buffer and subjected to western blot analysis using an anti-phosphorus-p44 / 42 MAP kinase antibody (ERK1 / 2) and an anti-ERK antibody

245 (Cell Signaling). The percentage of phosphorylated ERK versus total ERK was determined in this way.

In addition, the proliferation assay based on factor-dependent mouse BaF3 cells used frequently for cytokine receptors can also be developed and applied.

Among the hybridoma supernatants tested in the human FGF21 ELK-luciferase reporter assay based on CHO cells (clone 2E10), more than 30 were identified as positive (> 5% of FGF21 activity) when compared to 20 nN of FGF21 as positive control. Antibodies were then purified from the conditioned media of these positive hybridoma cultures and tested again in the CHO cell-based ELK-luciferase reporter assay. Figure 7 showed the representative antibodies in the dose-response potency assay with an estimated EC50 of less than 1 pg / ml (or 6.7 nM). The activities were confirmed in the ERK1 / 2 phosphorylation assay based on L6 cells (Figure 8) with EC50 of less than 10 nM, which is consistent with the ELKluciferase assay in the stable CHO 2E10 cell line.

EXAMPLE 6

FGF21 TYPE SIGNAL INDUCTION IS SPECIFIC TO THE FGFRlc / pKLOTHO COMPLEX

FGF21 has been reported to signal through multiple receptor complexes, including FGFRlc, 2c, 3c and 4, when paired with β-Klotho. The selectivity of FGF21 agonist antibodies was tested in rat L6 myeloblastic cells transfected with vectors that express the respective FGFRs and pKlotho. The results shown in

246

Figure 9 demonstrate that the activity was mediated selectively and exclusively by means of FGFRlc, and not by means of FGFR2c, 3c or 4 when they were paired with β-Klotho, as no activity was detected in the last receptors up to 100 nM of the agonist antibodies. This unique selectivity strongly suggests that the action of these antibodies is βKlotho-dependent, although it must specifically involve the FGFRlc component of the signaling complex.

EXAMPLE 7

ACTIVITY IN HUMAN PRIMARY ADIPOCYTES

FGF21 stimulates glucose uptake and lipolysis in cultured adipocytes, and adipocytes are considered to be more physiologically relevant than the recombinant reporter cell system. A panel of antibodies has been shown to exhibit FGF21-like Erk phosphorylation activity in the human adipocyte assay (Figure 10) with an EC50 estimated to be less than 10 nM.

EXAMPLE 8

COMPETITIVE CONNECTION AND EPITOP CONNECTION

To compare the similarity of antibody binding sites at the FGF21 receptor, a series of competitive binding experiments was performed and measured by Biacore. In one example (and as shown in Figure 11), two representative FGF21 receptor agonist antibodies (24H11 and 17D8) and antibodies binding to a non-functional FGF21 receptor (1A2.1) were immobilized on the surface of the sensor chip. Soluble human FGFRlc / β-Klotho ECD-Fc complex or β-Klotho was then captured on the surfaces of the immobilized antibody. Finally, several of the test FGF21 receptor antibodies were injected individually

247 on the captured human soluble FGF21 or β-Klotho receptor. If the injected antibody recognizes a distinct binding site from that recognized by the immobilized antibody, a second binding event will be observed. If the antibodies recognize a very similar binding site, binding will no longer be observed.

As shown in Figure 11A, there are two distinct, though partially overlapping, binding sites for the tested agonist antibodies. One site is covered by 24H11, 21H2, 18B11.1 and 17C3 (Group A) and the other site is covered by 17D8, 12E4 and 18G1 (Group B). The two non-functional antibodies, 2G10 and 1A2, bind to different sites between them and are distinct from the two sites covered by the agonist antibodies in Groups A and B. Other functional antibodies that bind to the Group A epitope included 20D4, 22H5, 16H7 , 40D2 and 46D11. It was demonstrated by this method that two other functional antibodies, 26H11 and 37D3, bind to the same site covered by Group B antibodies. In addition, a third binding site for functional antibodies has been identified for 39F11, 39F7 and 39G5 (group C) , which appeared to be distinct from the binding sites of Groups A and B (Figure 11B).

Another Biacore analysis was performed with biotinylated FGF21 immobilized on the sensor chip. Ten nM of soluble β-Klotho were then passed on the chip alone or mixed with the individual test antibodies at 100 nM. Figure 12 showed that several agonist antibodies in Group A (24H11, 18B11, 17C3) and antibody 12E4 (Group B) competed significantly with FGF21 for binding to soluble βKlotho, while non-functional antibodies 2G10

248 and 1A2 and several other functional antibodies showed no competitive link with FGF21.

Figure 11C summarizes the connection results obtained.

EXAMPLE 9 RECOGNITION OF NATIVE AND DENATURED STRUCTURES

The ability of revealed antigen-binding proteins to recognize denatured and native structures was investigated. The procedure and results were as follows.

Example 9.1

Agonist FGF21 receptor antibodies do not recognize denatured structures, as demonstrated by FACS

Lysates of cells from CHO cells stably expressing FGF21 receptor (FGFRlc and β-Klotho) or parental CHO cells were diluted with sample buffer without beta-mercaptoethanol (non-reducing conditions). Twenty μΐ of cell lysate were streaked on separate adjacent streaks with a molecular weight marker streak on 4-20% SDS-PAGE gels. After electrophoresis, the gels were blotted on 0.2 μ nitrocellulose filters. The blots were treated with saline solution buffered with Tris / Triton-X (TBST) plus 5% skimmed milk (blocking buffer) for 30 minutes. The blots were then cut along the molecular weight marker streaks. The strips were then probed with FGF21 receptor agonist antibodies (12C3, 26H11, 12E4, 21H2, 18B11 or 20D4), and murine antiβ-Klotho or anti-huFGFRl from commercial goat mice (R&D Diagnostics) in TBST / 5% milk . The blots were incubated with the antibodies for one hour at

249 environment, followed by three washes with TBST + 1% milk. The blots were then probed with secondary anti-human or goat-HRP antibodies for 20 min. The blots received three 15 min washes with TBST, followed by treatment with Pierce Supersignal West Dura development reagent (1 min) and exposure to the X-ray film Kodak Biomax.

The commercial anti-β-Klotho and anti-FGFR1 antibodies detected the corresponding receptor proteins in the SDSPAGE, indicating that they bind to the denatured receptor proteins. In contrast, none of the FGF21 receptor agonist antibodies tested detected the corresponding protein species, suggesting that they bind to the native conformational epitope distinct from commercial antibodies that bind to denatured sequences.

Example 9.2

Agonist FGF21 receptor antibodies bind to the native structure of the receptor, as demonstrated by FACS

A FACS binding assay was performed with several commercially available FGFRlc and β-Klotho antibodies, and with several of the revealed FGF21 receptor agonist antibodies. The experiments were done as follows.

CHO cells that stably express the FGF21 receptor were treated with R&D Systems mouse anti-huFGFRl, goat anti-mu β-Klotho or FGF21 receptor antibodies 24H11, 17C3, 17D8, 18G1 or 2G10 (1 pg by 1 x 10 ⁶ cells in 100 µl of PBS / 0.5% BSA). The cells were incubated with the antibodies at 4 ° C, followed by two washes with PBS / BSA. The cells were then treated with secondary antibodies labeled with FITC at 4 ° C, followed by two washes. The cells were resuspended in 1 ml of

250

PBS / BSA and antibody binding was analyzed using a FACS Calibur instrument.

Consistent with the western blot results, all FGF21 receptor agonist antibodies tested will bind well to the cell surface FGF21 receptor in FACS, whereas commercial anti-p-Klotho or anti-FGFR1 antibodies did not. This observation further confirmed that FGF21 receptor agonist antibodies recognize the native structure, whereas commercial antibodies to receptor components do not.

EXAMPLE 10

ARGININE SCAN

As described above, antigen binding proteins that bind to human FGF21R, for example, FGFRlc, β-Klotho or both FGFRlc and β-Klotho, were created and characterized. To determine the neutralizing determinants in human FGFRlc and / or β-Klotho that these various antigen binding proteins have bound to, several mutant FGFRlc and / or β-Klotho proteins that have arginine substitutions in selected amino acid residues can be constructed. FGFRlc and / or human β-Klotho. Arginine scanning is a recognized method of assessing where antibodies, or other proteins, bind to another protein; see, for example, Nanevicz et al., (1995) J. Biol. Chem., 270: 37, 21.619-21.625 and Zupnick et al., (2006) J. Biol. Chem., 281: 29, 20.46420.473. In general, the arginine side chain is positively charged and relatively bulky when compared to other amino acids, which can disrupt antibody binding to a region of the antigen where the mutation is

251 introduced. Arginine scanning is a method that determines whether a residue is part of a neutralizing determinant and / or an epitope.

Various amino acids distributed by the extracellular domains of human FGFRlc and / or β-Klotho can be selected for mutation in arginine. Selection can be directed to charged or polar amino acids to maximize the possibility of the residue being on the surface and reduce the likelihood of the mutation resulting in a wrongly folded protein. Using standard techniques known in the art, sense and antisense oligonucleotides that contain the mutated residues can be designed based on criteria provided by the Stratagene Quickchange® II protocol kit (Stratagene / Agilent, Santa Clara, CA). Mutagenesis of the FGFRlc and / or β-Klotho wild type (WT) sequences can be performed using a Quickchange® II kit (Stratagene). Chimeric constructs can be designed to encode a FLAG-histidine tag (six histidines (SEQ ID. NO: 382)) at the carboxy terminus of the extracellular domain to facilitate purification via the polyHis tag. Multiplex analysis using the workstation and BioPlex software (BioRad, Hercules, CA) can be performed to determine neutralizing determinants in human FGFRlc and / or βKlotho by analyzing the differential binding of exemplary human FGFRlc and / or β-Klotho mAbs to arginine mutants versus FGFRlc and / or wild-type β-Klotho proteins. Any number of pentaHis coated blood cell codes (penta-His revealed as SEQ ID. NO .: 383) (Qiagen, Valencia, CA; see wwwl.qiagen.com) can be used to capture protein with

252 histidine tag. Blood cell codes can allow multiplexing of FGFRlc and / or βKlotho and FGFRlc and / or wild-type human β-Klotho arginine mutants.

To prepare the globules, 100 μΐ of FGFRlc and / or β-Klotho supernatants of FGFRlc and / or β-Klotho transient expression culture are bound to globules coated with penta-His (pentaHis revealed as SEQ ID NO: 383) overnight at 4 ° C or 2 hours at room temperature with vigorous stirring. The globules are then washed as per the manufacturer's protocol and the pool of globules pooled and aliquoted into 2 or 3 columns of a 96-well filter plate (Millipore, Bellerica, MA, product # MSBVN1250) for test points in duplicate or triplicate, respectively. One hundred μΐ of antiFGFRlc and / or anti-β-Klotho antibodies in 4-fold dilutions are added to the wells, incubated for 1 hour at room temperature, and washed. One hundred μΐ of a 1: 100 dilution of PE-conjugated human IgG anti-Fc (Jackson Labs., Bar Harbor, ME, product # 109-116-170) is added to each well, incubated for 1 hour at room temperature and washed. The globules are resuspended in 1% BSA, shaken for 3 minutes, and read on a Bio-Plex workstation. The binding of the antibody to the arginine mutant of the FGFRlc and / or β-Klotho protein is compared to the binding of the antibody to human FGFRlc and / or wild-type β-Klotho from the same pool. An antibody titration over a scale of approximately one log can be performed. The median fluorescence intensity (MFI) of the FGFRlc and / or β-Klotho protein arginine mutant can be represented

253 graphically as a percentage of the maximum human wild-type FGFRlc and / or β-Klotho signal. Those mutants for which the signal for all antibodies are below a cutoff value, for example, 30% wild-type FGFRlc and / or β-Klotho, can be considered to have a very low protein concentration in the globule as a result low expression in the transient culture or possibly erroneous folding, and can be excluded from the analysis. Mutations (ie arginine substitutions) that increase the EC50 for FGFRlc and / or β-Klotho mAb by a cut-off value, for example, 3 times or more (as calculated, for example, by GraphPad Prism®) can be considered to have negatively affected the binding of FGFRlc and / or β-Klotho mAb. Through these methods, neutralizing determinants and epitopes for various antibodies to FGFRlc and / or β-Klotho are elucidated.

EXAMPLE 11

CONSTRUCTION OF CHEMICAL RECEIVERS

In another method of determining the activation determinants in human FGFRlc and / or β-Klotho to which these various antigen-binding proteins bind, chimeric FGFRlc and / or β-Klotho proteins can be constructed and tested among human species and mouse cells, expressed in transient or stable 293 or CHO cells, as described above. For example, a chimeric FGF21 receptor, which comprises FGFRlc, FGFR2C, FGFR3C or native human FGFR4, in an example FGFRlc, paired with human / mouse chimeric β-Klotho in which regions or sequences selected in β-Klotho human are systematically

254 replaced by the corresponding specific mouse residues (see, for example, Figure 2A-2C). Similarly, native human β-Klotho paired with human / mouse chimeric FGFRlc, FGFR2c, FGFR3c or FGFR4, in an example FGFRlc, in which regions or sequences selected in human FGFRlc are systematically replaced by the corresponding specific mouse residues (see, for example, by example, the alignments of Figures 1A-1B). The crucial sequences involved in the binding and / or activity of the antigen binding proteins can be derived by means of the binding assay or activity measures described in previous Examples 4, 5, 6 and 7 based on the chimeric FGF21 receptors.

Example 11.1

Construction of specific chimeras

Mouse human-β-Klotho chimeras were constructed using the methodology described in Example 14. A schematic representation of the constructed chimeras is shown in Figure 29; briefly, the generated chimeras were composed (from the N to the C terminal) by a fusion of a sequence of human β-Klotho fused to a sequence of murine β-Klotho fused to a sequence of human βKlotho. Human β-Klotho (SEQ ID. NO .: 8) was used as a framework in which regions of murine β-Klotho (full-length sequence shown in SEQ ID NO: 468) were inserted. The regions of murine β-Klotho that were inserted were as follows:

Murine waste 82P-520P

255

PKNFSWGVGTGAFQVEGSWKTDGRGPSIWDRYVYSHLRGVNGTDRSTDSYIFLEKDLL ALDFLGVSFYQFSrSWPRLFPNGTVAAVNAQGLRYYRALLDSLVLRNlEPTVTLYHWDLP LTLQEEYGGWKNATMlDLFNDYATYCFQTFGDRVKYWnHNPYLVAWHGFGTGMHA PGEKGNLTAVYTVGHNLIKAHSKVWHNYDKNFRPHQKGWLSITLGSHWIEPNRTDNM EDVINCQHSMSSVLGWFANP1HGDGDYPEFMKTGAMIPEFSEAEKEEVRGTADFFAFSF GPNNFRPSNTVVKMGQNVSLNLRQVLNWÍKLEYDDPQ1L1SENGWFTDSYIKTEDTTAIY MMKNFLNQVLQAIKFDEIRVFGYTAWTLLDGFEWQDAYTTRRGLFYVDFNSEQKERKP KSSAHYYKQIIQDNGFPLKESTPDMKGRFP (SEQ ID NO:.. 470)

Murine residues 506F-1043S

FPLKESTPDMKGRFPCDFSWGVTESVLKPEFTVSSPQFTDPHLYVWNVTGFJRLLYRVEG VRLKTRPSQCTDYVSIKKRVEMLAKMKVTHYQFALDWTSILPTGNLSKVNRQVLRYYR CWSEGLLY

DLVKLWITINEPNRLSDMYNRTSNDTYRAAHNLM1AHAQVWHLYDRQYRPVQHGAVS LSLHCDWAEPANPFVDSHWKAAERFLQFE1AWFADPLFKTGDYPSVMKEY1ASKNQRG LSSSVLPRFTAKESRLVKGTVDFYALNHFTTRFVIHKQLNTNRSVADRDVQFLQDITRLS SPSRLAVTPWGVRKLLAWIRRNYRDRDIYÍTANGIDDLALEDDQIRKYYLEKYVQEALK

AYL1DKVKIKGYYAFKLTEEKSKPRFGFFTSDFRAKSSVQFYSKLISSSGLPAENRSPACG QPAEDTDCTICSFLVEKKPLIFFGCCF1STLAVLLSJTVFHHQKRRKLFQKARNLQNIPLKK N ° DE.

Murine residues 1M-193L

MKTGCAAGSPGNEWIFFSSDERNTRSRKTMSNRALQRSAVLSAFVLLRAVTGFSGDGK AlWDKKQYVSPVNPSQLFLYDTFPKNFSWGVGrGAFQVEGSWKTDGRGPSIWDRYVYS HLRGVNGTDRSTDSYIFLEKDLLALDFLGVSFYQFSlSWPRLFPNGTVAAVNAQGLRYY RALLDSLVLRN1EPIVTL (SEQ ID NO:.. 472)

Murine waste 82P-302S

PKNFSWGVGTGAFQVEGSWKTDGRGPSJWDRYVYSHLRGVNGTDRSTDSYIFLEKDLL ALDFLGVSFYQFSISWPRLFPNGTVAAVNAQGLRYYRALLDSLVLRNIEPIVTLYHWDLP LTLQEEYGG WKNATMIDLFN DYAT YCTQTFGDRVKY W1TIHNP LVAWHGFGTGMHA PGEKGNLTAVYTVGHNL1KAHSKVWHNYDKNFRPHQKGWLS1TEGS Y (SEQ ID NO:.. 473)

Murine residues 194Y-416G

YHWDLPLTLQEEYGGWKNATMIDLFNDYATYCFQTFGDRVKYWIT1HNPYLVAWHGF GTGMHAPGEKGNLTAVYTVGHNLIKAHSKVWHNYDKNFRPHQKGWLSITLGSHWIEP NRTDNMEDVrNCQHSMSSVLGWFANPIHGDGDYPEFMKTGAMIPEFSEAEKEEVRGTA DFFAFSFGPNNFRPSNTVVKMGQNVSLNLRQVLNWIKLEYDDPQILISENG (SEQ ID NO:.. 474)

Murine residues 302S-506F

256

S H W1EPN RTDNM ED VINCQHSMSS VLGWFANPIHGDGD YPEFMKTGAMIPEFSEAEKEE VRGTADFFAFSFGPNNFRPSNTVVKMGQNVSLNI.RQVLNWTKEEYDDPQlLlSENGWFr DSYIKTED'ITAIYMMKNFLNQVLQAIKFDEIRVFGY I'AW 1 LLDGFEWQDAY ΓΊ RRGLFY VDFNSEQKERKPKSSAHYYKQIIQDNGF (DESEQ ID NO:.. 475)

Murid waste 416G-519P

GWFTDSYIKTEDTTAIYMMKNFLNQVLQAJKFDEIRVFGYTAWTLLDGFEWQDAYTTR RGLFYVDFNSEQKERKPKSSAHYYKQIIQDNGFPLKESTPDMKGRF (SEQ ID: 476)

PLKESTPDMK.GRFPCDFSWGVTESVLKPEFTVSSPQFTDPHLYVWNVTGNRLLYRVEGV RLKTRPSQCTDY VS1 KKRVEMLAKMKVrHYQFALD WTSILPTGNLSKVNRQ VLR YYRC VQSGLL.

Murine waste 520P-735A

PCDFSWGVTESVI.KPEFTVSSPQFTDPFILYVWNVTGNRI.LYRVEGVRI.KTRPSQCTDY VSTKKRVEMLAKMKVTHYQFALDWTSILPTGNLSKVNRQVLRYYRCVVSEGLK1.GVFP MVTLYHPTHSUI.GLPLPLLSSGGWLNMNTAKAFQDYAEJ.CFRELGDLVKLWITÍNF.PNR LSDMYNRTSNDTYRAAHNLMIAHAQVWHLYDRQYRPVQHGA (SEQ ID NO:.. 478)

Murine residues 632G-849Q

GVFPMVrLYHPrHSFlLGLPLPLLSSGGWLNMNlAKAFQDYAELCFRELGDLVKLWrnN EPNRLSDMYNRTSNDTYRAAHNLMIAHAQVWHLYDRQYRPVQHGAVSLSLHCDWAE PANTFVDSHWKAAERFLQFEIAWFADPLFKTGDYPSVMKEYIASKNQRGLSSSVLPRFT AKESRLVKGTVDFYALNHFTTRFVIHKQLNTNRSVADRDVQFLQ (SEQ ID NO:.. 479)

Murine residues 735A-963S

AVSI.SI.IICDWAEPANPFVDSHWKAAERFLQFEIAWFADPI.FKTGDYPSVMKEYIASKN QRGl.SSSVLPRFTAKESRLVKGTVDFYALNHFTTRFVÍHKQLNIKRSVADRDVQFLQDJ 1 'RLSSPSRLAVTPWGVRKLLAW1RRNYRDRD1YITANGIDDLALEDDQ1RKYYLEKYVQE ALKAYLIDKVKIKGYYAFKLTEEKSKPRFGFFTSDFRAKSSVQFYSKLJSSS (SEQ ID NO:.. 480)

Murine residues 1M-81F

MKTGCAAGSPGNEWIFFSSDERNTRSRKTMSNRALQRSAVLSArVLLRAVTGFSGDGK AIWDKKQYVSPVNPSQLFLYDTF (SEQ ID. NO: 481)

257

Murine waste 82P-193L

PKNFSWGVGTGAFQVEGSVVKTDGRGPSiWDRYVYSIlLRGVNGTDRSTDSYIFLEKDLL

ALDFLGVSFYQFSISWPRLFPNGTVAAVNAQGLRYYRALLDSLVLRNIEPIVTL (SEQ ID. NO: 482)

The chimeras generated using the murine β-Klotho sequences comprised the following components:

258

C-terminal residues of human βKlotho 523-1,044 194-1,044 303-1,044 419-1,044 509-1,044 522-1,044 635-1,044 Mouse β-Klotho residues 82-520 506-1,043 1-193 82-302 194-416 302-506 416-519 507-632 N-terminal residues of human βKlotho 00 1-507 00 1 1-193 1-301 1-417 1-508 ID No. SEQ. of construction Construction identifier huBeta_Klotho (1-81, 523-1.044) (muBetaKLOTHO 82-520) huBeta_Klotho (1-507) (muBetaKLOTHO S06F-1045S) huBeta_Klotho (194-1,044) (muBetaKLOTHO 1-L193) huBeta_Klotho (1-81, 303- 1,044) (muBetaKLOTHO 82P-302S) huBeta_Klotho (1-193, 419- 1.044) (muBetaKLOTHO Y194-416G) huBeta_Klotho (1-301, 509- 1.044) (muBetaKLOTHO S302-F506) huBeta_Klotho (1-417, 522- 1.044) (muBetaKLOTHO G416-F519) huBeta_Klotho (1-507, 635-

259

C-terminal residues of human βKlotho 738-1,044 852-1,044 967-1,044 82-1,044 194-1,044 Mouse β-Klotho residues 520-735 632-849 735-963 CO 1 82-193 N-terminal residues of human βKlotho 1-521 1-633 1-736 00 1 ID No. SEQ. of construction Construction identifier 1.044) (muLeta KLOTHO F06-G632) huBeta_Klotho (1-521, 738- 1.044) (KLOTHO 520P-735A muBeta) huBeta_Klotho (1-633, 852- 1,044) (muLeta KLOTHO 632G-849Q) huBeta_Klotho (1-736, 967- 1,044) (muLeta KLOTHO 735A-963S) huBeta_Klotho (82-1,044) (muBeta KLOTHO 1-81F) huBeta_Klotho (1-81,194-1,044) (muBeta KLOTHO 82P-193L)

260

The generated chimeras comprised the following amino acid sequences:

(i) huBeta_Klotho (1-81, 523-1.044) (muBetaKLOTHO 82-

520)

MKPGCAAGSPGNEWIFFSTDElTTRYRNTMSNGGLQRSViLSALILLRAV TGFSGDGRAIWSKNPNFTPVNESQLFLYDTFPKNFSWGVGTGAFQVEGSW KTDGRGPSIWDRYVYSHLRGVNGTDRSTDSYIFLEKDLLALDFLGVSFYQ FSISWPRLFPNGTVAAVNAQGLRYYRALLDSLVLRNIEPIVTLYHWDLPL TLQEEYGGWKNATMIDLFNDYATYCFQTFGDRVKYWITIHNPYLVAWHGF GTGMHAPGEKGNLTAWTVGHNLIKAHSKVWHNYDKNFRPHQKGWLSrrL GSHWIEPNRTDNMEDVINCQHSMSSVLGWFANPIHGDGDYPEFMKTGAM1 PEFSEAEKEEVRGTADFFAFSFGPNNFRPSNTVVKMGQNVSLNLRQVLNW IKLEYDDPQILISENGWFTDSYIKTEDTTAIYMMKNFLNQVLQAIKFDET RVFGYTAWTLLDGFEWQDAYTTRRGLFYVDFNSEQKERKPKSSAHYYKQI rQDNGFPLKESTPDMKGRFPCDFSWGVTESVLKPESVASSPQFSDPHLYV WNATGNRLLHRVEGVRLKTRPAQCTDFVNIKKQLEMLARMKVTHYRFALD WASVLPTGNLSAVNRQALRYYRCVVSEGLKLGISAMVTLYYPTHAHLGLP EPLLHADGWLNPSTAEAFQAYAGLCFQELGDLVKLWlTINEPNRLSDiYN RSGNDTYGAAHNLLVAHALAWRLYDQQFRPSQRGAVSLSLHADWAEPANP YADSHWRAAERFLQFEJAWFAEPLFKTGDYPAAMREYIASKHRRGLSSSA LPRLTEAERRLLKGTVDFCALNHFTTRFVMHEQLAGSRYDSDRDIQFLQD ITRLSSPI'RLAVIPWGVRKLLRWVRRNYGDMDIYITASGIDDQALEDDRL RKYYLGKYLQEVLKAYLIDKVRIKGYYAFKI.AEEKSKPRFGFFTSDFKAK SSlQFYNKVlSSRGFPFENSSSRCSQTQ ENTECrVCLFLVQKKPLIFLGC CFFSTLVLLLSIAJFQRQKRRKFWKAKNLQHIPLKKGKRWS (ID. SEQ. N °: 455) (ii) huBeta_Klotho (1-507) (muBetaKLOTHO S06F-1.045S)

MKPGCAAGSPGNEWIFFSTDEITTRYRNTMSNGGLQRSVILSALILLRAV

TGFSGDGRAIWSKNPNFTPVNESQLFLYDTFPKNFFWGIGTGALQVEGSW

KKDGKGPSIWDHFIHTHLKNVSSTNGSSDSYIFLEKDLSALDFIGVSFYQ FSlSWPRLFPDGr / TVANAKGLQYYSTLLDALVLRNIEPTVTLYHWDLPL ALQEKYGGWKNDTIIDIFNDY ATYCFQMFGDRVKY WIT1HN PY L V WHGY GTGMHAPGEKGNLAAVYTVGHNLIKAHSKVWHNYNTHFRPHQKGWLSrTL GSFIWIEPNRSENTMDIFKCQQSMVSVLGWFANPIHGDGDYPEGMRKKLFS VI.PIFSEAEKHEMRGTADFFAFSFGPNNFKPLNTMAKMGQNVSLNLREAL NW1KLEYNNPRILIAENGWFTDSRVKTEDTTATYMMKNFLSQVLQAIRLD EiRVFGYTAWSLLDGFEWQDAYTIRRGLFYVDFNSKQKERKPKSSAHYYK

QllRENGFPLKESTPDMKGRFPCDFSWGVTESVLKPEFTVSSPQFTDPHL YVWNVTGNRLLYRVEGVRLKTRPSQCTDYVSIKKRVEMLAKMKVTHYQFA LDWTSILPTGNLSKVNRQVLRYYRCVVSEGLKLGVFPMVTLYHPTFISHLG .ΕΡΕΡΕΕ880αψΕΝΜ'ΝΤΑΚΑ.Ρ9ΟΥΑΕΕΕΕΚΕΕ00ΕνΚΕ'ΨΙΤΙΝΕΡΝΚΕ8ΟΜ YNRTSNDTYRAAFrNLMIAHAQVWHLYDRQYRPVQHGAVSLSLHCDWAEPA

261

NPFVDSHWKAAERFLQFEIAWFADPLFKTGDYPSVMKEYIASKNQRGLSS SVLPRFTAKESRLVKGTVDFYALNHFTrRFVIHKQLNTNRSVADRDVQFL QDITRLSSPSRLAVTPWGVRKLLAWIRRNYRDRDIYITANGJDDLALEDD QÍRKYYLEKYVQEALKAYL1DKVKIKGYYAFKLTEEKSKPRFGFFTSDFR AKSSVQFYSKLISSSGLPAENRSPAGGQPAEDTDCTICSFLVEKXPLIFF GCCF1STLAVLLS1TVFHHQKRRKFQKARNLQN1PLKKGHSRVFS (SEQ ID NO:.. 456) (iii) huBeta_Klotho (194-1044) (muBetaKLOTHOl-L193) MKTGCAAGSPGNEWIFFSSDERNTRSRKTMSNRALQRSAVLSAFVLLRAV TGFSGDGKAIWDKKQYVSPVNPSQLFLYDTFPKNFSWGVGTGAFQVEGSW KTDGRGPSIWDRYVYSHLRGVNGTDRSTDSY1FLEKDLLALDFLGVSFYQ FS1SWPRLFPNGTVAAVNAQGLRYYRALLDSLVLRN1EPIVTLYHWDLPL ALQEKYGGWKNDTIIDIFNDYATYCFQMFGDRVKYWmHNPYLVAWHGY GTGMHAPGEKGNLAAWTVGHNLIKAHSKVWHNYNTHFRPHQKGWLSITL GSHWIEPNRSENTMD1FKCQQSMVSVLGWFANP1HGDGDYPEGMRKKLFS VLPIFSEAEKHEMRGTADFFAFSFGPNNFKPLNTMAKMGQNVSLNLREAL NWIKLEYNNPRILIAENGWFTDSRVKTEDTTAIYMMKNFLSQVLQAIRLD E1RVFGYTAWSLLDGFEWQDAYTIRRGLFYVDFNSKQKERKPKSSAHYYK Q11RENGFSLKESTPDVQGQFPCDFSWGVTESVLKPESVASSPQFSDPHL YVWNATGNRXLHRVEGVRLKTRPAQCTDFVNIKKQLEMLARMKVTHYRFA LD WAS VLFTGN LSAVN RQALRY YRCVVSEGLKLGISAMVTLYYPTHAHLG LPEPLLHADGWLNPSTAEAFQAYAGLCFQELGDLVKLWITINEPNRLSDI YNRSGNDTYGAAHNLLVAHALAWRLYDQQFRPSQRGAVSLSLHADWAEPA NPYADSHWRAAERFLQFEIAWFAEPLFKTGDYPAAMREYIASKHRRGLSS SALPRLTEAERRLLKGTVDFCALNHFrj'RFVMHEQLAGSRYDSDRDlQFL QDFFRLSSPTRLA VIP WGVRKLLR WVRRN Y GDMD1Y1TASGÍDDQAL EDD RLRKYYLGKYLQEVLKAYL1DKVR1KGYYAFKLAEEKSKPRFGFFTSDFK AKSSrQFYNKVlSSRGFPFENSSSRCSQTQENTECTVCLFLVQKKPLIFL GCCFFSTLVLLLSIAIFQRQKRRKFWAKNLQHIPLKKGKRVVS (id. SEQ. N °: 457) (iv) huBeta_Klotho (1-81, 303-1.044) (muBetaKLOTHO

82P-302S)

MKPGCAAGSPGNFAVIFFSTDEITTRYRNTMSNOGLQRSVILSAI.ILLRAV TGFSGDGRAIWSKNPNFTPVNESQLFLYDTFPKNFSWGVGTGAFQVEGSW KTDGRGPSJ WDR Y VYSHLRGVNGTDRSTDSYJFLEKDLLALDFLGVSFYQ FSISWPRLFPNGTVAAVNAQGLRYYRALLDSLVLRNIEPIVTLYHWDLPL TLQEEYGGWKNATMIDLFNDYATYCFQTFGDRVKYWlTfflNPYLVAWHGF GTGMHAPGEKGNLTAVYTVGHNLIKAHSKVWHNYDKNFRPHQKGWLSITL GSHWIEPNRSENTMDIFKCQQSMVSVLGWFANPIHGDGDYPEGMRKKLFS VLPIFSEAEKHEMRGTADFFAFSFGPNNFKPLNTMAKMGQNVSLNLREAL NWIKLEYNNPRILIAENGWFTDSRVKTEDTTAIYMMKNFLSQVLQAIRLD EIRVFGYTAWSLLDGFEWQDAYTIRRGLFYVDFNSKQKERKPKSSAHYYK qiirengfslkestpdvqgqfpcdfswgvtesvlkpesvasspqfsdphl

262

YVWNATGNRLLHRVEGVRLKTRPAQCTDFVNIKKQLEMLARMKVTHYRFA LDWASVLPTGNLSAVNRQALRYYRCVVSEGLKLGISAMVTLYYPTHAHLG LPEPLLHADGWLNPSTAEAFQAYAGLCFQELGDLVKL \ MT [NEPNRLSDI YNRSGNDTYGAAHNLLVAHALAWLYDQQFRPSQRGAVSLSLHADWAEPA NPYADSHWRAAERFLQFEIAWFAEPLFKTGDYPAAMREYIASKHRRGLSS SALPRLTEAERRLLKGTVDFCALNHFTTRFVMHEQLAGSRYDSDRD1QFL QDITRLSSPTRLAVIPWGVRKLLRWVRRNYGDMDIYITASGIDDQALEDD RLRKYYLGKYLQEVLKAYLÍDKVRIKGYYAFKLAEEKSKPRFGFFTSDFK AKSSIQFYNKV1SSRGFPFENSSSRCSQTQENTECTVCLFEVQKKPLIFL GCCFFSTLVLLLSIAIFQRQKRRKFWKAKN LQH1PLKKGKRWS (SEQ ID NO:.. 458) (v) huBeta_Klotho (1-193, 419-1044) (muBetaKLOTHO

Y194-416G)

MKPGCAAGSPGNEWIFFSTDEITTRYRNTMSNGGLQRSVILSAULLRAV TGFSGDGRAIWSKNPNFTPVNESQLFLYDTFPKNFFWGIGTGALQVEGSW KKDGKGPSlWDHFIIlTIlLKNVSSrNGSSDSYlFLEKDLSAEDFIGVSFYQ FSIS WPRLFPDGIVTV ANAKGLQYYSTLLDA L VLRNIE PIVTL YHWDLPL _{TLQEEYGG. W} ^ _ATMIDL p _NDYATYCF Q _{TFGDRVKYWrnHNpYLVAWHGF} GTGMHAPGEKFiNLTAVYTVGIINLIKAHSKVWHNYDKNFRPHQKGWLSlTL. GSHWIEPNRTDNMEDVINCQHSMSSVLGWFANPIHGDGDYPEFMKTGAMI PEFSEAEKEEVRGTADFFAFSFGPNNFRPSNTVVKMGQNVSLNLRQVLNW IKLEYDDPQlIJSENGWFTDSRVKTEDTTAIYMMKNFLSQVLQAIRLDEr RVFGYTAWSLLDGFEWQDAYTÍRRGLFYVDFNSKQKERKPKSSAHYYKQI TRENGFSLKESTPDVQGQFPCDFSWGVTESVLKPESVASSPQFSDPHLYV WNATGNRLLHRVEGVRLKTRPAQCTDFVNIKKQLEMLARMKVTHYRFALD WASVLPTGNLSAVNRQALRYYRCVVSEGLKLG1SAMVTLYYPTHAHLGLP NIRS-HADGWLNPSTAEAFQAYAGLCFQELGDLVKI.WITINEPNRLSDIYN rsgndtygaahnllvahalawrlydqqfrpsqrgavslslhadwaepanp YADSHWRAAERFLQFE1AWFAEPLFKTGDYPAAMREY1ASKHRRGLSSSA LPRLTEAERRLLKGTVDFCALNHFTTRFVMHEQLAGSRYDSDRDIQFLQD ITRLSSPTRLAVIPWGVRKLLRWVRRNYGDMDIYJTASGIDDQALEDDRL · RKYYLGKYLQEVLKAYLIDKVRIKGYYAFKLAEEKSKPRFGFFTSDFKAK SS1QFYNKV1SSRGFPFENSSSRCSQTQENTECTVCLFLVQKKPL1FLGC CFFSTLVLLLSIAIFQRQKRRKFWKAKNLQHTPLKKGKRVVS (SEQ ID NO:.. 459) (vi) huBeta_Klotho (1-301, 509-1044) (muBetaKLOTHO

S302-F506)

MKPGCAAGSPGNEWIFFSTDEITTRYRNTMSNGGLQRSVILSALILLRAV TGFSGDGRA1WSKNPNFTPVNESQLFLYDTFPKNFFWGIGTGALQVEGSW KKDGKGPSIWDHFIHTHLKNVSSTNGSSDSY1FLEK.DLSALDFIGVSFYQ FS1SWPRLFPDGIV TVANAKGLQY YSTLLDALVLRNIEPIVTLYHWDLPL ALQEKYGGWKNDTIlDIFNDYATYCFQMFGDRVKYWmHNPYLVAWHGY

263

GTGMHAPGEKGNLAAVYTVGHNLIKAHSKVWHNYNTHFRPHQKGWLSITL GSHWIEPNRTDNMEDVINCQHSMSSVLGWFANPIHGDGDYPEFMKTGAMI PEFSEAEKEEVRGTADFFAFSFGPNNFRPSNTWKMGQNVSLNLRQVLNW IKLEYDDPQILISENGWFTDSYIKTEDTTAIYMMKNFLNQVLQAIKFDEl RVFGYTAWTLLDGFEWQDAYTTRRG LFY VDFNSEQKERKPKSSAHYYKQ1 IQDNGFSLKESTPDVQGQFPCDFSWGVTESVLKPESVASSPQFSDPHLYV WNATGNRLLHRVEGVRLK'I'RPAQCTDFVNIKKQLEMLARMKVTHYRFALD WAS VLPTGN LS VN RQ ALRYYRCVVSEGLKLGIS AM VTLYYPTHAHLGLP EPLLIIADGWLNPSTAEAFQAYAGLCFQELGDLVKLW1TINEPNRLSDIYN RSGNDTYGAAHNLLVAHALAWRLYDQQFRPSQRGAVSLSLHADWAEPANP YADSHWRAAERFLQFE1AWFAEPLFKTGDYPAAMREY1ASKHRRGLSSSA LPRLTEA ERRLLKGTVDFCA LN HFTTRF VM H EQ LAGSR YDSDRDIQFLQD ITRLSSPTRLAVIPWGVRKLLRWVRRNYGDMDIYITASGIDDQALEDDRL RKYYLGKYLQEVLKAYLIDKVRIKGYYAFKLLAEEKSKPRFGFFTSDFKAK SSIQFYNKVISSRGFPFENSSSRCSQTQENTECTVCLFLVQKKPLIFLGC C'FFSTLAIJISIAIFQRQKRRKFWKAKNEQll IPI.KKGKRVVS (ID. SEQ. N °: 460) (vii) huBeta_Klotho (1-417, 522-1.044) (muBetaKLOTHO G416-F519)

MKPGCAAGSPGNEWIFFSTDEITTRYRNTMSNGGLQRSVILSALILLRAV TGFSGDGRAIWSKNPNFTPVNESQLFLYDTFPKNFFWGIGTGALQVEGSW KKDGKGPSIWDHFIHTHLKNVSSTNGSSDSYIFLEKDLSALDFIGVSFYQ FS1SWPRLFPDG1VTVANAKGLQYYSTLLDALVLRN1EPIVTLYHWDLPL ALQEK.YGGWKNDTnDIFNDYATYCFQMFGDRVKYWrriHNPYLVAWHGY GTGMHAPGEKGNLAAVYTVGHNLIKAHSKVWHNYNTHFRPHQKGWLSÍTL GSHW1EPNRSENTMD1FKCQQSMVSVLGWFANP1HGDGDYPEGMRKKLFS VLPIFSEAEKHEMRGTADFFAFSFGPNNFKPLNTMAKMGQNVSLNLREAL NWIKLEYNNPRILIAENGWFTDSYIKTEDTTAIYMMKNFLNQVLQAIKFD EIRVFGYTAWTLLDGFE WQDAYTTRRGLFY VDFN SEQKERKPKSS HY YK QIIQDNGFPLKESTPDMKGRFPCDFSWGVTESVLKPESVASSPQFSDPHL YVWNATGNRLLHRVEGVRLKTRPAQCTDFVNIKKQLEMLARMKVTHYRFA LDWASVLPTGNLSAWRQALRYYRCWSEGLKLGISAMVTLYYPTHAHLG LPEPLLHADGWLNPSTAEAFQAYAGLCFQELGDLVKLWT1NEPNRLSDI YNRSGNDn'GAAHNLLVAHALAWRLYDQQFRPSQRGAVSLSLHADWAEPA NPYADSHWRAAERFLQFELAWFAEPLFKTGDYPAAMREYIASKHRRGLSS SALPRLTEAERRLLKGTVDFCALNHFTrRFVMHEQLAGSRYDSDRDIQFL QDΓΓRLSSPTRLAVlPWGVRKLL · RWVRRNYGDMDlYIΐASGIDDQALEDD RLRKYYLGKYLQEVLKAYLIDKVRIKGYYAFKLAEEKSKPRFGFFTSDFK AKSSIQFYNKVISSRGFPFENS SSRCSQTQENTECTVCLHWQKKPLIFL GCCFFSTLVLLLSIAIFQRQKRRKFWKAKNLQinPLKKGKRVVS (ID. SEQ. N °: 461) (viii) huBeta_Klotho (1-507, 635-1.044) (muBeta KLOTHO

F06-G632)

264

MKPGCAAGSPGNEWJFFSTDEmRYRNTMSNGGLQRSVJLSALTLLRAV TGFSGDGRAIWSKNPNFTPVNESQLFLYDTFPKNFFWGIGTGALQVEGSW KKDGKGPSIWDHFIHTHLKNVSSTNGSSDSYIFLEKDLSALDFIGVSFYQ FSISWPRLFPDGIVTVANAKGLQYYSTLLDALVLRNIEPJVTLYHWDLPL ALQEKYGGWKNDTODIFNDYATYCFQMFGDRVKYWmHNPYLVAWHGY GTGMHAPGEKGNLAAVYTVGHNL1KAHSKVWHNYNTHFRPHQKGWLSITL GSHWIEPNRSENTMD1FKCQQSMVSVLGWFANP1HGDGDYPEGMRKFLLFS VLPIFSEAEKIIEMRGTADFFAFSFGPNNFKPLNTMAKMGQNVSLNLREAl. NWIKLEWNPRILLAENGWFTDSRVKTEDTTAIYMMKNFLSQVLQAIRLD E1RVFGYTAWSLLDGFEWQDAYTIRRGLFYVDFNSKQKERKPKSSAHYYK QIIRENGFPI.KESTPDMKGRFPCDFSWGVTESVLKPEFTVSSPQFTDPIIL YVWNVTGNRI.IARVEGVRLKTRPSQCTDYVSIKKRVEMEAKMKVTHYQFA lOWTSILPTGNLSKVNRQVLRYYRCVVSEGLKLGlSAMVTLYYPTHAHLG LPEPLLHADGWLNPSTAEAFQAYAGLCFQELGDLVKLWlTfNEPNRLSDI YNRSGNDTYGAAHNLLVAFIALAWRI.YDQQFRPSQRGAVSLSLHADWAEP / X NPYADSHWRAAERFLQFEÍAWFAEPLFK.TGDYPAAMREY1ASKHRRGLSS SALPRLTEAERRLLKGTVDFCALNHFTTRFVMHEQLAGSRYDSDRDIQFL QD1TRLSSPTRLAV IP WGVRKLLLR WVRRN Y GDMD1YITASGIDDQ ALEDD RLRKYYLGkYLQEVLKAYLIDKVRlKGYYAFKLAEEKSKPRFGFFTSDFK AKSS1QFYNKVISSRGFPFENSSSRCSQTQENTECTVCLFLVQKKPL1FL GCCFFSTLVLLLSIAIFQRQKRRKFWKAKNLQHIPLKKGKRVVS (SEQ ID NO:.. 462) (ix) huBeta_Klotho (1-521, 738 -1,044) (muLeta KLOTHO

520P-735A)

MKPGCAAGSPGNEWIFFSTDEnTRYRNTMSNGGLQRSVILSALTLLRAV TGFSGDGRAIWSKNPNFTPVNESQLFLYDTFPKNFFWGIGTGALQVEGSW KKDGKGPSIWDHFIHTHLKNVSSTNGSSDSYIFLEKDLSALDFIGVSFYQ FSISWPRLFPDGIVTVANAKGLQYYSTLLDALVLRNIEPIVTLYHWDLPL ALQEKYGGWKNDT1ID1FNDYATYCFQMFGDRVKYWITIHNPYLVAWHGY GTGMHAPGEKGNLAAVYTVGHNL1KAHSKVWHNYNTHFRPHQKGWLSITL GSHWlEPNRSEWMDiFKCQQSMVSVEGWFANPinGDGDYPEGMRKKLFS VLPIFSEAEKHEMRGTADFFAFSFGPNNFKPLNTMAKMGQNVSLNLREAL NWIKLEYNNPRILIAENG WFTDSRVKTEDTTAIYM M KN FLSQVLQAIRLD EIRVFGYTAWSIJ.DGFEWQDAYTIRRGLFYVDFNSKQKERKPKSSAHYYK QIIRENGFSLKESTPDVQGQFPCDFSWGVTESVLKPEFTVSSPQFTDPFIL YVWNVTGNRLLYRVEGVRLKTRPSQCTDYVSIKKRVEM1.AIGMKVTHYQFA IOWTSILPTGNESKVNRQVLRYYRCWSEGLKLGVFPMVTLYHPTHSHLG [, PEPI [. SSGGW1.NMNTAKAFQDYAELCFRELGDI.VKLWITINEPNR1.SDM YNRTSNDTYRAAHNLMIAHAQVWHLYDRQYRPVQHGAVSLSLHADWAEPA NPYADSHWRAAERFLQFEIAWFAEPLFKTGDYPAAMREY1ASKHRRGLSS SALPRLTEAERRLLKGTVDFCALNHFrrRFVMHEQLAGSRYDSDRDlQFL QDrrRLSSPIRLAVIPWGVRKLLRWVRRNYGDMDIYITASGlDDQ ALEDD RLRKYYLGKYLQEVLKAYLlDKVRIKGYYAFKLAEEKSKPRFGFFrSDFK AkSSIQFYNKVISSRGFPF ENSSSRCSQTQENTECTVCLFLVQKKPLIFL GCCFFSTLVLLLSIAIFQRQKRRKFWKAKNLQinPLKKGKRVVS

265 (SEQ ID NO: 463) (x) huBeta_Klotho (1-633, 852-1.044) (muBeta KLOTHO

632G-849Q)

MKPGCAAGSPGNEW1FFSTDEITTRYRNTMSNGGLQRSVILSALILLRAV TGFSGDGRAIWSKNPNFTPVNESQLFLYDTFPKNFFWGIGTGALQVEGSW KKDGKGPSIWDFIFIHTHLKNVSSTNGSSDSYII'LEKDL.SAI.DFIGVSFYQ FSL · SWPRLFPDGlVTVANAKGLQYYSTLLDALVLRNlEPlVTLYHWDLPL ALQEKYGGWKNDTIIDIFNDYATYCFQMFGDRVKYWITIHNPYLVAWHGY GTGMHAPGEKGNLAAVYTVGHNLIKAHSKVWHNYNTHFRPHQKGWLSITL GSHWIEPNRSENTMDIFKCQQSMVSVLGWFANP1HGDGDYPÊGMRKKLFS VLPIFSEAEKHEMRGTADFFAFSFGPNNFICPLNTMAKMGQNVSLNLREAL NWIKLEYNNPRILIAENGWFTDSRVKTEDTTA1YMMKNFLSQVLQAIRLD EIRVFGYTAWSLLDGFEWQDAYTIRRGLFYVDFNSKQKERKPKSSAHYYK QIIRENGFSLKESTPDVQGQFPCDFSWGVTESVLKPESVASSPQFSDPUL YVWNATGNRLLHRVEGVRLKTRPAQCTDFVNIKKQLEMLARMKVTHYRFA LDWASVLPTG.NLSAVNRQALRYYRCVVSEGLKLGVFPMVTLYHPTHS.HLG LPLELLSSGGWLNMNTAKAFQDYAELCFRELGDLVKLW1TINEPNRLSDM YNRTSNDTYRAAHNLMIAHAQVWHLYDRQYRPVQHGAVSLSLHCDWAEPA NPFVDSFIWKAAERFLQFEIAWFADPLFKTGDYPSVMKEYIASKNQRGLSS SVLPRFrAKESRLVKGTVDFYALNHFTl'RFVIHKQLNTNRSVADRDVQFL QD1TRLSSPTRLAVIPWGVRKLLRWVRRNYGDMD1YITASG1DDQALEDD RLRKYYLGKYLQEVLKAYLIDKVRJKGYYAFKLAEEKSKPRFGFFTSDFK AKSSIQFYNKVISSRGFPF ENSSSRCSQTQENTECTVCLFLVQKKPLIFL GCCFFSTLVLLLSIAIFQRQKRRKFWKAKNLQHIPLKKGKRVVS (ID. SEQ. N °: 464) (xi) huBeta_Klotho (1-736, 967-1.044) (muBeta KLOTHO 735A-963S)

MKPGCAAGSPGNEWIFFSTDErn’RYRNTMSNGGLQRSVILSALILLRAV

TGFSGDGRAIWSKNPNFTPVNESQLFLYDTFPKNFFWGIGTGALQVEGSW KKDGKGPSIWDHFIHTHLKNVSSTNGSSDSYJFLEKDLSALDF1GVSFYQ FSISWPRLFPDGIVTVANAKGLQWSTLLDALVLRNfEPTVTLYHWDLPL ALQEKYGGWKNDniDIFNDYATYCFQMFGDRVKYWrnHNPYLVAWHGY GTGMHAPGEKGNLAAVYTVGHNLIKAHSKVWHNYNTHFRPHQKGWLS1TL GSHW1EPNRSENTMD1FKCQQSMVSVLGWFANP1HGDGDYPEGMRKKLFS VLPIFSEAEKHEMRGTADFFAFSFGPNNFKPLNTMAKMGQN ^T VSLNLREAL nwikleynnpriliaengwftdsrvktedttaiymmknflsqvlqairld E1RVFG ΥΤΛ WSI I .DGFE WQDAYTIRRGLFYVDFN S YK KQKERKPKSS ΑΗΥ QIIRENGFSLKESTPDVQGQFPCDFSWGVTESVLKPESVASSPQFSDPFTL yvwnatgnrllhrvegvrlktrpaqctdfvnikkqlemlarmkvthyrfa ldwasvlptgnlsavnrqalryyrcvvseglklgisamvtlyypthahlg lpepllhadgwlnpstaeafqayaglcfqelgdlvklwitinepnrlsdi ynrsgndtygaahnllvabalawrlydqqfrpsqrgavslslhcdwaepa npfvdshwkaaerflqfeiawfadplfktgdypsvmkeyiasknqrglss SVLPRFTAK £ SRLVKGTVDFYALNFlFTrRFVliIKQLNTNRSVADRDVQFL

266

QDITRLSSPSRLA VTPWG VRKLLA W1RRNY RDRDIY1TANGIDDLA LE DD QIRKYYLEKYVQEALKAYL1DKVK1KGYYAFKLTEEKSKPRFGFFTSDFR AKSSVQFYSKLISSSGFPFENSSSRCSQTQENTECTVCLFLVQKKPLIFL GCCFFSTLVLLLSIATFQRQKRRKFWKAKNLQHIPLKKGKRVVS (SEQ ID NO:.. 465) (xii) huBeta_Klotho (82-1044) (1-81F muBeta KLOTHO)

MKTGCAAGSPGNEWIFFSSDERNTRSRKTMSNRALQRSAVLSAFVl.LRAV TGFSGDGKAfWDKKQYVSPVNPSQLFLYDTFPKNFFWGIGTGALQVEGSW KKDGKGPSJ WDH FIHTH G KN VSSTNGSSDSY1FLEKDLSALDFIG VSFYQ PSISWPRLFPDGIVTVANAKGLQYYSTLLDALVLRNJEPIVTLYHWDLPL ALQEKYGGWKNDTIIDIFNDYATYCFQMFGDRVKYWITIHNPYLVAWHGY GTGMHAPGEKGNLAAX ^ YTVGHNLIKAHSKVWHNYNTHFRPHQKGWLSITL GSHWLEPNRSENTMDIFKCQQSMVSVLGWFANP1HGDGDYPEGMRKKLFS VLPIFSEAEKÍ1EMRGTADFFAFSFGPNNFKPLNTMAKMGQNVSLNLREAL NWIKLEYWPRILIAENGWFTDSRVKTEDTTAFYMMKNFLSQVLQAIRLD EIRVFGYTAWSLLDGFEWQDAYTIRRGLFYVDFNSKQKERKPKSSAHYYK QIIRENGFSLKESTPDVQGQFPCDFSWGVTESVLKPESVASSPQFSDPHL YVWNATGNRLLHRVEGVRLKTRPAQCTDFVNJKKQLEMLARMKVTHYRFA LDWASVLPTGNLSAVNRQALRYYRCWSEGLKLGISAMVTLYYPTHAHLG LPEPLLHADGWLNPSTAEAFQAYAGLCFQELGDLVKLWITINEPNRLSDI YNRSGNDTYGAAHNLLVAHALAWRLYDQQFRPSQRGAVSLSLHADWAEPA NPYADSHWRAAERFLQFEIAWFAEPLFKTGDYPAAMREYIASKHRRGLSS SALPRLTEAERRLLKGTVDFCALNHFITRFVMHEQLAGSRYDSDRDIQFL QDITRLSSPTRLAVIPWGVRKLLRWVRRNYGDMDIYITASGIDDQALEDD RLRKYYLGKYLQEVLKAYLIDKVRJKGYYAFKLAEEKSKPRFGFFTSDFK AKSSIQFYNKVISSRGFPFENSS SRCSQTQENTECTVCLFLVQKKI’LIFL GCCFFSTLVLLLSlAIFQRQKRRKLFWkAKNLQHIPLKKGKRVVS (ID. SEQ. N °: 466) (xiii) huBeta_Klotho (1-81, 194-1,044) (muBeta KLOTHO 82P-193L)

MKPGCAAGSPGNEWIFFSTDEITTRYRNTMSNGGLQRSVILSALTLLRAY ^ TGFSGDGRAIWSKNPNFTPVNESQLFLYDTFPKNFSWGVGTGAFQVEGSW KTDGRGPSIWDRYVYSHLRGVNGTDRSTDSYIFLEKDLLALDFLGVSFYQ FSISWPRLFPNGTVAAVNAQGLRYYRALLDSLVLRNIEPIVTLYHWDLPL ALQEKYGG WKNDT1IDIFNDYATYCFQ MFGDRVKY witih N PYLV The WHGY gtgmhapgekgnlaavytvghnlikahskvwhnynthfrphqkgwlsitl GSHW1EPNRSENTMD1FKCQQSMVSVLGWFANP1HGDGDYPEGMRKKLFS VLPIFSEAEKHEMRGTADFFAFSFGPNNFKPLNTMAKMGQNVSLNLREAL NW1KLEYNNPR1LIAENGWFTDSRVKTEDTTA1YMMKNFLSQVLQA1RLD EIRVFGYTAWSLÍ.OGFEWQDAYTJRRGLFYVDFNSKQKERKPKSSAHYYK QIIRENGFSLKESTPDVQGQFPCDFSWGVTESVLKPESVASSPQFSDPHL YVWNATGNRLLHRVEGVRLKTRPAQCTDFVNIKKQLEMLARMKVTHYRFA

267

LDWASVLPTGNLSAVNRQALRYYRCVVSEGLKL.GISAMVTLYYPTHAHLG LPEPLLHADGWLNPSTAEAFQAYAGLCFQELGDLVKLWITINEPNRLSDI YNRSGNDTYGAAHNLLV HA ALA WR LYDQQFRPSQRG VS LSLHADWAEPA NPYADSHWRAAERFLQFEIAWFAEPLFKTGDYPAAMREYIASKBRRGLSS SALPRLTEAERRLLKGTVDFCALNHFTTRFVMHEQLAGSRYDSDRDIQFL QDITRLSSIGRLAVIIÃVG WKLL.RW VRRNYGDMDIYITASGIDDQALEDD RLRKYYLGKYLQEVLKAYLIDKVRIKGYYAFKLAEEKSKPRFGFFTSDFK AKSSIQFYNKVISSRGFPFENSSSRCSQTQENTECTVCLFLVQKKPLIFL GCCFFSTLVLLLSIAIFQRQKRRKFWKAKNLQHTPLKKGKRVVS (SEQ ID NO:.. 467)

Several antigen-binding proteins provided here, as well as human FGF21, have been tested for their ability to activate chimeras in L6 cells. Figure 30 correlates the results observed with each molecule tested.

These data indicate that while human FGF21 was able to activate FGFRlc combined with all human / mouse β-Klotho chimeras (the + sign indicates receptor activity), mouse sequence substitutions in human β-Klotho affected the activities of 16H7, 37D3 and 39F7. See Figure 30. These results suggest that β-Klotho sequences 1-81, 302-522 and 849-1.044 are important for the activities of agonist antigen binding proteins and may represent an important epitope for their function.

EXAMPLE 12

PROTEASE PROTECTION ANALYSIS

Regions of the human FGF21 receptor bound by antigen binding proteins that bind to the human FGF21 receptor, for example, FGFRlc, β-Klotho or FGFRlc and β-Klotho complex, can be identified by fragmentation of the human FGF21 receptor into peptides with specific proteases, for example, AspN, Lys-C,

268 chymotrypsin or trypsin. The sequence of the resulting human FGF21 receptor peptides (i.e., peptide fragments that both contain disulfide and disulfide from portions of FGFRlc and β-Klotho) can then be determined. In one example, soluble forms of a human FGF21 receptor, for example, a complex comprising the FGFRlc ECD-Fc heterodimer and β-Klotho ECD-Fc described herein, can be digested with AspN (which cleaves after residues of aspartic acid and some glutamic acid at the amino end) by incubating about 100 pg of FGF21 receptor soluble at 1.0 mg / ml in 0.1 M sodium phosphate (pH 6.5) for 20 hours at 37 ° C with 2 pg of AspN.

A peptide profile of AspN digestion can then be generated in HPLC chromatography, while a control digest with a similar amount of antibody is expected to be essentially resistant to AspN endoprotease. A protease protection assay can then be performed to determine the proteolytic digestion of the human FGF21 receptor in the presence of the antigen binding proteins. The general principle of this assay is that the binding of an antigen-binding protein to the FGF21 receptor can result in protection of certain specific protease dividing sites and that information can be used to determine the region or portion of the FGF21 receptor where the antigen binding protein binds.

Briefly, the peptide digestion for being submitted to the peptide mapping by HPLC; individual peaks are collected, and peptides are identified and mapped by LC-MS analysis by online electrospray ionization (ESI-LC-MS) and / or by sequencing the

269 terminal N. HPLC analysis for these studies can be performed using a narrow hole reverse phase C18 column (Agilent Technologies) for offline analysis and use of a reverse phase 5 capillary C18 column (the separation group) for LC- MS. HPLC peptide mapping can be performed with a linear gradient from 0.05% trifluoroacetic acid (mobile phase A) to 90% acetonitrile in 0.05% trifluoroacetic acid. Columns can be developed at a desired flow rate for 10 narrow-hole HPLC for offline or online LC-MS analysis, and for capillary HPLC for online LC-MS analysis.

Sequence analyzes can be performed by LC-MS / MS online and by Edman sequencing on the 15 peptide peaks recovered by HPLC. ESI LC-MS online analyzes of peptide digestion can be performed to determine the precise mass and sequence of the peptides that are separated by HPLC. The identities of selected peptides present in the peptide peaks of the 20 protease digest can thus be determined.

EXAMPLE 13

CYNOMOLGUS MONKEY STUDY

A construct encoding the antigen-binding protein designated herein 16H7 was generated using the methodology disclosed in Examples 1-3. 16H7 was expressed, purified and characterized as described in Examples 1-5 and was studied in vivo in obese Cynomolgus monkeys. 16H7 is a fully human IgGl antibody and is described by the sequences provided in Tables 1-4, supra.

0 Example 13.1

270

Study design study was carried out on obese Cynomolgus monkeys.

The monkeys were 8-19 years old. Their body weights ranged from 7-14 kg and the BMI ranged from 36-74 kg / m ² . The monkeys were acclimated for 6 weeks before starting to administer the compound. During the acclimation period, monkeys were familiarized with study-related procedures, including restraint in chairs, subcutaneous injection (PBS, 0.1 ml / kg), oral fattening (water, 10 ml / kg) and blood drawn for samples. non-OGTT and OGTT. After 4 weeks of training, OGTT and baseline plasma metabolic parameters were measured. Twenty monkeys were selected and randomized into two treatment groups to obtain similar baseline levels of body weight, OGTT glucose profiles and blood glucose and triglyceride levels.

study was carried out blindly. Vehicle (n = 10), 16H7 (n = 10). The compound was given every other week (5 mg / kg). In the week when animals were not injected with 16H7, they were given vehicle injection instead. After 2 injections of 16H7, the animals were monitored for an additional 6 weeks to eliminate the compound and recover from the treatments. Feed intake, body weight, clinical biochemistry and OGTT were monitored throughout the study. Feed intake was measured at each meal. Body weight was measured weekly. Blood samples were collected on different days in a fasted or prandial state to measure glucose, insulin and triglyceride levels. OGTTs were performed every two weeks after the study started. 0 day

271 when treatment starts is designated 0 and ο detailed study planning is shown in Figure 14.

The results presented in this Example represent data collected over the 68 days of the study.

Example 13.2

Effect of 16H7 on feed intake

The animals were fed twice a day, with each animal receiving 120 g of formulated feed established during the acclimation period. The remaining feed was removed and weighed after each meal to calculate feed intake. The feeding periods were from 8 am until 8:30 am (± 30 minutes) and after 4:30 pm until 5:00 pm (± 30 minutes). Fruits (150 g) were provided to each animal from 11:30 am to 12:30 pm (± 30 minutes) every day.

Compared to vehicle, 16H7 reduced feed intake in monkeys. The effect decreased and the feed intake returned close to baseline or control levels after about 21 days of treatment. 16H7 had no significant effect on feed intake in the morning (Figure 15) and only modestly reduced feed intake in the afternoon meal during treatment (Figure 16). An increase in feed intake in the morning was observed after the 49th day (Figure 15). Throughout the study (and even during the acclimation period), fruit intake appeared to be lower in the 16H7 group compared to the vehicle group. Overall, 16H7 showed a significant effect on the inhibition of feed intake.

Example 13.3

272

Effect of 16H7 on body weight

Body weight was monitored weekly throughout the study. Throughout the 4-week treatments, the body weight of animals treated with vehicle remained constant, while the body weight of animals treated with 16H7 decreased progressively. Body weight did not return to baseline at the end of the 6-week elimination period (Figure 17).

Example 13.4

16H7 effect on body mass index (BMI), waist circumference (AC) and skinfold thickness (FTS)

BMI, AC and SFT were monitored weekly throughout the study, both pre- and post-administration of test compound when body weight was measured. BMI is defined as the individual's body weight divided by the square of his height. SFT is the thickness of a double layer of skin and the fat below it is measured with a compass. BMI, SFT and AC are relatively accurate, simple and inexpensive measurements of body composition, particularly indicative of subcutaneous fat. Vehicle-treated animals showed relatively stable BMI, SFT and AC throughout the study. Animals treated with 16H7 showed decreased levels of BMI, AC and FTS over the 4-week study period, suggesting that the 16H7 compound resulted in reduced fat mass. The results are shown in Figures 18-20, respectively. These measured parameters did not return to baseline values at the end of the 6-week elimination period.

Example 13.5

273

Effect of 16H7 on the oral glucose tolerance test (OGTT)

OGTTs were performed before and after the start of treatments. Before 16H7 injections, baseline values for glucose and insulin levels were measured over the OGTT (Figures 21 and 22, respectively) and were not statistically significantly different between the vehicle and 16H7 groups. Post-dose OGTTs were performed every two weeks during the treatment period and after 3 weeks of the elimination period. 16H7 slightly increased glucose tolerance after 4 weeks of treatment and 3 weeks of elimination period. The animal model used is not glucose-intolerant, which explains the modest effects observed (Figure 21). Insulin levels were statistically significantly decreased in animals treated with 16H7 (significance observed at time 0 during OGTT performed after 2 weeks of treatment, at time 0 and 15 minutes during OGTT performed after 4 weeks of treatment and at time 0 and 60 minutes during OGTT performed after 2 weeks of treatment) (Figure 22).

Example 13.6

16H7 effect on fasting and prandial glycemia and insulin levels

Blood was collected from animals fasting overnight or in prandial conditions after feeding in the morning. Under fasting conditions, blood was drawn weekly 5 days after each injection. In prandial conditions, blood was taken on the 2nd, 11th, 16th, 25th and 46th days after the first injection. 16H7

274 did not reduce fasting or prandial blood glucose levels (Figures 23 and 25). No hypoglycemia was observed in any of the monkeys treated with 16H7. 16H7, however, resulted in a statistically significant decrease in plasma fasting and insulin levels (Figures 24 and 26).

Example 13.7

Effect of 16H7 on triglyceride levels

Measurements were made of the same samples collected for glucose and insulin measurements. Triglyceride levels were significantly reduced in animals treated with 16H7 when measured under fasting or prandial conditions (Figures 27 and 28).

Example 13 · 8

Conclusions

In a study of obese male Cynomolgus monkeys, animals treated with 16H7 showed improved metabolic parameters. Body weight was reduced and body composition was improved. Short-term reduction in feed intake was observed and the effect decreased and feed intake recovered at baseline or control levels with 21 days in the study. The levels of fasting insulin and triglycerides were also reduced by 16H7. Insulin levels measured during OGTT have also been improved.

EXAMPLE 14

VARIANT FORMS OF ANTIGEN BINDING PROTEINS 16H7 AND 22H5

The 16H7 and 22H5 antigen binding proteins, which are described here in Tables 1-4, have been mutated to

275 transmit different properties to the molecule, for example, changes in solubility, pl, global charge, immunogenicity in humans and animal models, stability etc. The mutations were composed of additions, deletions or substitutions in the light chain (designated LC, SEQ ID NO: 14) or in the heavy chain (designated HC, SEQ ID NO: 32) of the molecule. The single point mutations revealed were made individually or two or more mutations were combined.

Examples of mutations and combinations of mutations that were introduced into the 16H7 heavy and light chain sequences include the following:

I83K (in the 16H7 heavy chain) (SEQ ID. NO: 396)

E16Q (on the 16H7 heavy chain) + V24F (on the 16H7 heavy chain) + I83T (on the 16H7 heavy chain) + S100I (on the 16H7 heavy chain) + T119L (on the 16H7 heavy chain) (SEQ ID. N °: 395) D109S (in the 16H7 heavy chain) (SEQ ID. N °: 401)

Deletion of Y107 (in the 16H7 heavy chain) (SEQ ID. N °: 400)

Insertion of a Y residue on the N-terminal side of Y107 (in the 16H7 heavy chain) (SEQ ID. NO: 405)

D88R + P89A + V90E (in the 16H7 heavy chain) (ID. DE

SEQ. N °: 398) D4 9Y (at jail Light in 16H7) (ID . IN SEQ. N °: 386) D4 9A (at jail Light in 16H7) (ID . IN SEQ. N °: 387) D91A (at jail Light in 16H7) (ID . IN SEQ. N °: 388) D4 9A (at jail Light in 16H7) + D91A (at light chain 16H7) (ID. SEQ. N °: 389) Q16K (at jail Light in 16H7) (ID . IN SEQ. N °: 385)

276

Examples of mutations and combinations of mutations that were introduced into the 22H5 heavy and light chain sequences include the following:

N92Q (in the 22H5 light chain) (SEQ ID. NO: 402)

S94A (in the 22H5 light chain) (SEQ ID. NO: 403)

C109S (in the 22H5 heavy chain) (SEQ ID. NO: 404)

Briefly, the generated antigen binding proteins comprised the following 16H7 heavy and light chain pairs:

(i) 16H7 light chain (ID. SEQ. N °: 14) paired with an 16H7 heavy chain what comprises I83K (ID. OF SEQ. N °: 396); (ii) 16H7 light chain (ID. SEQ. N °: 14) paired with an 16ΗΊ heavy chain i who understands E16Q , V24F, I83T, S100I, T119L (SEQ ID. . N °: : 3 95); (iii) 16H7 light chain (ID. . SEQ. N °: : 14) paired with an 16H7 heavy chain what comprises D109S (ID. OF SEQ. N °: 401); (iv ) 16H7 light chain (ID. SEQ. N °: 14) paired with an 16H7 heavy chain what comprises the deletion of Y107 (ID . SEQ. No.: 400); (v) 16H7 light chain (ID. SEQ. N °: 14) paired with an 16H7 heavy chain what understands the i insertion of

a Y residue on the N-terminal side of Y107 (SEQ ID. NO: 405);

(vi) 16H7 light chain (SEQ ID NO: 14) paired with a 16H7 heavy chain comprising D88R, P89A, V90E (SEQ ID NO: 398);

(vii) 16H7 heavy chain (SEQ ID NO: 32) paired with a 16H7 light chain comprising D49Y

277 (SEQ ID. NO: 386);

(viii) 16H7 heavy chain (SEQ ID NO: 32) paired with a 16H7 light chain comprising D4 9A (LC) (SEQ ID NO: 387);

(xi) 16H7 heavy chain (SEQ ID NO: 32) paired with a 16H7 light chain comprising D91A (SEQ ID NO: 388);

(ix) 16H7 heavy chain (SEQ ID NO: 32) paired with a 16H7 light chain comprising D4 9A, D91A (SEQ ID NO: 389);

(x) 16H7 heavy chain (SEQ ID NO: 32) paired with a 16H7 light chain comprising Q16K (LC) (SEQ ID NO: 3 85);

and the following pairs of 22H5 heavy and light chain sequences:

(xi) 22H5 heavy chain (SEQ ID NO: 31) paired with a 22H5 light chain comprising N92Q (LC) (SEQ ID NO: 402);

(xii) 22H5 heavy chain (SEQ ID NO: 31) paired with a 22H5 light chain comprising S94A

(LC) (SEQ ID. NO: 403); (xiii) 22H5 chain lightweight (SEQ ID NO: 13) paired with a heavy chain 22H5 which comprises C109S (HC) (ID. SEQ. No. 4 04); (xiv) 22H5 chain lightweight (SEQ ID NO: 13) paired with a 22H5 heavy chain comprising a insertion

of a tyrosine residue at position 107 (SEQ ID NO: 405).

The amino acid sequences for the generated light chain variants are shown in Table 6:

278

Table 6A - Amino acid sequences of 16H7 and 22H5 variants

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279

Μ ο Q S Q Hi Η Μ ω 387 385 Variant chain amino acid sequence TECS SYVLTQPPSVSVAPGQTARITCGGNNIGSESVHWY QQKPGQAPVLWYADSDRPSGIPERFSGSNSGNTA TLTISRVEAGDEADYYCQVWDGNSDHWFGGGTKL TVLGQPKANPTVTLFPPSSEELQANKATLVCLISD FYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYA ASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP TECS SYVLTQPPSVSVAPGKTARITCGGNNIGSESVHWY QQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTA TLT IS RVEAGDEAD YYCQVWDGNSDHWFGGGTKL TVLGQPKANPTVTLFPPSSEELQANKATLVCLISD FYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYA ASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP ID No. SEQ. of the paired sequence 32 32 Paired with H3 H3 Variation σι Q O Center string 16H7 light chain 16H7 light chain

280

κ Q K Q Ò Η W ω CD 00 388 Variant chain amino acid sequence TECS SYVLTQPPSVSVAPGQTARITCGGNNIGSESVHWY QQKPGQAPVLWYYDSDRPSGIPERFSGSNSGNTA TLTISRVEAGDEADYYCQVWDGNSDHWFGGGTKL TVLGQPKANPTVTLFPPSSEELQANKATLVCLISD FYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYA ASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP TECS SYVLTQPPSVSVAPGQTARITCGGNNIGSESVHWY QQKPGQAPVLWYDDSDRPSGIPERFSGSNSGNTA TLTISRVEAGDEADYYCQVWAGNSDHWFGGGTKL TVLGQ PKANPTVTLFPPS SEELQANKATLVCLISD FYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYA ASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP ID No. SEQ. of the paired sequence CN m 32 Paired with m tn H3 Variation 05 + Q D91A Center string 16H7 light chain 16H7 light chain

281

ID. SEQ. N °: LO 00 co 386 Variant chain amino acid sequence TECS SYVLTQPPSVSVAPGKTARITCGGNNIGSESVHWY QQKPGQAPVLWYDDSDRPSGIPERFSGSNSGNTA TLTIS RVEAGDEADYYCQVWDGNSDHWFGGGTKL TVLGQPKANPTVTLFPPSSEELQANKATLVCLISD FYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYA ASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP TECS SYVLTQPPSVSVAPGQTARITCGGNNIGSESVHWY QQKPGQAPVLWYYDSDRPSGIPERFSGSNSGNTA TLTISRVEAGDEADYYCQVWDGNSDHWFGGGTKL TVLGQPKANPTVTLFPPSSEELQANKATLVCLISD FYPGAVTVAWKADGS PVKAGVETTKPSKQSNNKYA ASSYLSLTPEQWKSHRSYSCQTH ID No. SEQ. of the paired sequence CN CO 32 Paired with CO K H3 Variation _ w O D4 9Y Center string 16H7 light chain 16H7 light chain

282

ID. IN SEQ. N °: i__________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________ σι 00 co 385 Variant chain amino acid sequence TECS SYVLTQPPSVSVAPGQTARITCGGNNIGSESVHWY QQKPGQAPVLWYADSDRPSGIPERFSGSNSGNTA TLTISRVEAGDEADYYCQVWAGNSDHWFGGGTKL TVLGQPKANPTVTLFPPSSEELQANKATLVCLISD FYPGAVTVAWKADGS PVKAGVETTKPS KQSNNKYA ASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP TECS SYVLTQPPSVSVAPGKTARITCGGNNIGSESVHWY QQKPGQAPVLWYDDSDRPSGIPERFSGSNSGNTA TLTISRVEAGDEADYYCQVWDGNSDHWFGGGTKL TVLGQPKANPTVTLFPPSSEELQANKATLVCLISD FYPGAVTVAWKADGS PVKAGVETTKPS KQSNNKYA ASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP ID No. SEQ. of the paired sequence CN CO 32 Paired with ! co H3 Variation H σι Q + σ Q <0 O Center string 16H7 light chain 16H7 light chain

283

Μ o ’* Q! 3 Q C> Η M ω 00 m 390 Variant chain amino acid sequence TECS SYVLTQPPSVSVAPGQTARITCGGNNIGSESVHWY QQKPGQAPVLWYYDSDRPSGIPERFSGSNSGNTA TLTISRVEAGDEADYYCQVWDGNSDHWFGGGTKL TVLGQPKANPTVTLFPPSSEELQANKATLVCLISD FYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYA ASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP TECS QVTLKESGPVLVKPTETLTLTCTFSGFSLNNARMG VSWIRQPPGKALEWLAHIFSNDEKSYSTSLKSRLT IS KDTS KS Q WLIMTNMDPVDTAT YYCARSWTGG YYYDGMDVWGQGTTVTVSSASTKGPSVFPLAPCSR STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSWTVPSSNFGTQTYTCNVD ID No. SEQ. of the paired sequence CN ro 14 Paired with co L3 Variation ΟΪ ’φ Q V24F Center string 16H7 light chain 16H7 heavy chain

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296

ID. SEQ. N °: LT) 00 CO 386 Variant chain amino acid sequence QQGNVFSCSVMHEALHNHYTQKSLSLSPGK SYVLTQPPSVSVAPGKTARITCGGNNIGSESVHWY QQKPGQAPVLWYDDSDRPSGIPERFSGSNSGNTA TLTIS RVEAGDEAD YYCQVWDGNSDHWFGGGTKL TVLGQPKANPTVTLFPPSSEELQANKATLVCLISD FYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYA ASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP TECS SYVLTQPPSVSVAPGQTARITCGGNNIGSESVHWY QQKPGQAPVLWYYDSDRPSGIPERFSGSNSGNTA TLT ISRVEAGDEAD YYCQVWDGNSDHWFGGGTKL TVLGQPKANPTVTLFPPSSEELQANKATLVCLISD FYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYA ASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP ID No. SEQ. of the paired sequence σι co 32 Paired with co K H3 Variation ex D4 9Y Center string 16H7 light chain 16H7 light chain

297

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298

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299

Μ ο ”Q £ Q Ο »Η Μ ω 398 385 Variant chain amino acid sequence QVTLKESGPVLVKPTETLTLTCTVSGFSLNNARMG VSWIRQPPGKALEWLAHIFSNDEKSYSTSLKSRLT ISKDTSKSQWLIMTNMRAEDTATYYCARSWTGG YYYDGMDVWGQGTTVTVSSASTKGPSVFPLAPCSR STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSWTVPSSNFGTQTYTCNVD HKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFL FPPKPKDTLMISRTPEVTCWVDVSHEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTFRWSVLTWHQD WLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQ VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK SYVLTQPPSVSVAPGKTARITCGGNNIGSESVHWY ID No. SEQ. of the paired sequence 32 Paired with m H3 Variation D88R + P89A + V90E LO O Center string 16H7 heavy chain Jail

300

ID. IN SEQ. N °: k £> 00 CO 399 < h4 Q < PU X < Q Q PU 0 Ή Eh C0 The Eh co X (D s H H > The s Eh H > Pi Ό O 0 Q g Eh > 0 0 Q g Eh < <0 co 0 U g CO CO 0 0 s g u s 0 > co M M s 0 > co w co Q Q O > co co fc Q O > Q (d 0 > Eh Eh 0 0 > Eh Eh co Ό co > ω CO H co > co CO [χ < you W s £ U 0 g PL4 π £ PU 0 O QC Q W K s Pi Q w M co 0 M co < H M 0 M co Eh > 1 Ό PU 2 O H Fh 0 PU The O H Eh Eh Φ 1—1 O Q M > 0 | - | 0 l-l W > U u 4J 0 Ql M > HI Eh 0 Ql M > O Eh Aflj CO The M O O ] - { CO The M 0 u Q Q PU > co co Pi PU > CO co Eh £ pí HI co X Pi O co X h4 d Q O PU > CO Eh Q u PU > co Fh g co X PU PU Pi HI co X PU PU Pi Wl The s Q > 1 Ü4 co X O Q X co Eh Ql Q Q 0 CO PU XI Q Q 0 co PU (D X Eh Q < Eh Q Ό > W > The > > w The > > Q Eh O CO > Q Eh O Q (d HY 0 PU The M > Q 0 PU The w > ^to H > < The PU co > § < PU PU Q PU M > H PU PU M > Eh 0 > s Eh Q PU Ç > £ Eh H-l CO «D CX Pi PU CO O O Pi PU > CO M 0 co CX Q Eh 0 CO O < h4 W u ¹ pu | - | 0 0 X CO Q PU H 0 0 X co Q (1) Eh Q PU CO u > w H H-l PU CO u Eh W O Q > > 1 CO M > 1 O H-l > X CO w > O Eh Eh ÍJ-I <; Η co O Eh Eh <; Eh O d gives <d H <d Η u Ό ο ο Hi M ω d <0) <d 0) P <d CN CO H % IN 0) <n ASS <d d <d s CO CO <υ P 0 ü h4 (d ass + 0 CTi i <d o · X 00 ass <d ΟΊ •H P Q <d> 00 00 Q (d H rH 0) (D u <d TS Ί3 d «u = d P d 0) > Ή kO 0) Ό Φ > <D 0) τι 6 ¹ φ 0) the O 1) u CO

301

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ID No. SEQ. of the paired sequence 32 Paired with m

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302

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ID. SEQ. N °: Variant chain amino acid sequence HTFPAVLQSSGLYSLSSWTVPSSNFGTQTYTCNV DHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVF LFPPKPKDTLMISRTPEVTCVWDVSHEDPEVQFN WYVDGVEVHNAKTKPREEQFNSTFRWSVLTWHQ DWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREP QVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK ID No. SEQ. of the paired sequence Paired with Variation Center string

313

Table 6B

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 light chain Q16K TCCTATGTGCTGACTCAGCCACCCTCGGTGT CAGTGGCCCCAGGAAAGACGGCCAGGATT ACCTGTGGGGGAAACAACATTGGAAGTGA aagtgtgcactggtaccagcagaagccagg ccaggcccctgtgctggtcgtcfatgatga tagcgaccggccctcagggatccctgagcg attctctggctccaactctgggaacacggc caccctgaccatcagcagggtcgaagccgg ggatgaggccgactattactgtcaggtgtg ggatggtaatagtgaccatgtggtattcgg cggagggaccaagctgaccgtcctaggtca gcccaaggccaaccccactgtcactctgtt cccgccctcctctgaggagctccaagccaa caaggccacactagtgtgtctgatcagtga cttctacccgggagctgtgacagtggcctg gaaggcagatggcagccccgtcaaggcgg gagtggagaccaccaaaccctccaaacag agcaacaacaagtacgcggccagcagcta cctgagcctgacgcccgagcagtggaagtc ccacagaagctacagctgccaggtcacgca tgaagggagcaccgtggagaagacagtgg CCCCTACAGAATGTTCA 406

314

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 light chain D4 9Y TCCTATGTGCTGACTCAGCCACCCTCGGTGT CAGTGGCCCCAGGACAGACGGCCAGGATTA CCTGTGGGGGAAACAACATTGGAAGTGAA agtgtgcactggtaccagcagaagccaggc CAGGCCCCTGTGCTGGTCGTCTATTATGAT agcgaccggccctcagggatccctgagcga TTCTCTGGCTCCAACTCTGGGAACACGGCC ACCCTGACCATCAGCAGGGTCGAAGCCGGG GATGAGGCCGACTATTACTGTCAGGTGTGG GATGGTAATAGTGACCATGTGGTATTCGOC GGAGGGACCAAGCTGACCGTCCTAGGTCAG CCCAAGGCCAACCCCACTGTCACTCTGTTC CCGCCCTCCTCTGAGGAGCTCCAAGCCAAC AAGGCCACACTAGTGTGTCTGATCAGTGAC TTCTACCCGGGAGCTGTGACAGTGGCCTGG AAGGCAGATGGCAGCCCCGTCAAGGCGGG AGTGGAGACCACCAAACCCTCCAAACAGA GCAACAACAAGTACGCGGCCAGCAGCTACC TGAGCCTGACGCCCGAGCAGTGGAAGTCCC ACAGAAGCTACAGCTGCCAGGTCACGCATG AAGGGAGCACCGTGGAGAAGACAGTGGCC CCTACAGAATGTTCA 407

315

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 light chain D4 9A TCCTATGTGCTGACTCAGCCACCCTCGGTGT CAGTGGCCCCAGGAÇAGACGGCCAGGATTA CCTGTGGGGGAAACAACATTGGAAGTGAA AGTGTGCACTGGTACCAGCAGAAGCCAGGC CAGGCCCCTGTGCTGGTCGTCTATGCTGAT AGCGACCGGCCCTCAGGGATCCCTGAGCGA TTCTCTGGCTCCAACTCTGGGAACACGGCC ACCCTGACCATCAGCAGGGTCGAAGCCGGG GATGAGGCCGACTATTACTGTCAGGTGTGG GATGGTAATAGTGACCATGTGGTATTCGGC GGAGGGACCAAGCTGACCGTCCTAGGTCAG CCCAAGGCCAACCCCACTGTCACTCTGTTC CCGCCCTCCTCTGAGGAGCTCCAAGCCAAC AAGGCCACACTAGTGTGTCTGATCAGTGAC TTCTACCCGGGAGCTGTGACAGTGGCCTGG AAGGCAGATGGCAGCCCCGTCAAGGCGGG AGTGGAGACCACCAAACCCTCCAAACAGA GCAACAACAAGTACGCGGCCAGCAGCTACC TGAGCCTGACGCCCGAGCAGTGGAAGTCCC ACAGAAGCTACAGCTGCCAGGTCACGCATG AAGGGAGCACCGTGGAGAAGACAGTGGCC CCTACAGAATGTTCA 408

316

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 light chain D91A tcctatgtgctgactcagccaccgtcggtgt CAGTGGCCCCAGGACAGACGGCCAGGATTA CCTGTGGGGGAAACAACATTGGAAGTGAA AGTGTGCACTGGTACCAGCAGAAGCCAGGC CAGGCCCCTGTGCTGGTCGTCTATfíATGAT AGCGACCGGCCCTCAGGGATCCCrGAGCGA TTCTCTGGCTCCAACTCTGGGAACACGGCC ACCCTGACCATCAGCAGGGTCGAAGCCGGG GATGAGGCCGACTATTACTGTCAGGTGTGG GCTGGTAATAGTGACCATGTGGTATTCGGC GGAGGGACCAAGCTGACCGTCCTAGGTCAG CCCAAGGCCAACCCCACTGTCACTCTGTrC CCGCCCTCCTCTGAGGAGCTCCAAGC'CAAC AAGGCCACACTAGTGTGTCTGATCAGTGAC TTCTACCCGGGAGCTGTGACAGTGGCCTGG AAGGCAGATGGCAGCCCCGTCAAGGCGGG AGTGGAGACCACCAAACCCTCCAAACAGA GCAACAACAAGTACGCGGCCAGCAGCTACC TGAGÇCTGACGCCCGAGCAGTGGAAGTCCC ACAGAAGCTACAGCTGCCAGGTCACGCATG AAGGGAGCACCGTGGAGAAGACAGTGGCC CCTACAGAATGTTCA 409

317

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 light chain D4 9A + D91A TCCTATGTGCTGACTCAGCCACCCTCGGTGT CAGTGGCCCCAGGACAGACGGCCAGGATTA CCTGTGGGGGAAACAACATFGGAAGTGAA AGTGTGCACTGGTACCAGCAGAAGCCAGGC CAGGCCCCTGTGCTGGTCGTCTATGCTGAT AGCGACCGGCCCTCAGGGATCCCTGAGCGA _TTCTCTGGCTCC ^ _{CTCTGGGAACACGGCC} ACCCTGACCATCAGCAGGGTCGAAGCCGGG GATGAGGCCGACTATTACTGTCAGGTGTGG GCTGGTAATAGTGACCATGTGGTATTCGGC GGAGGGACCAAGCTGACCGTCCTAGGTCAG CCCAAGGCCAACCCCACTGTCACTCTGTTC CCGCCCTCCTCTGAGGAGCTCCAAGCCAAC AAGGCCACACTAGTGTGTCTGATCAGTGAC TTCTACCCGGGAGCTGTGACAGTGGCCTGG AAGGCAGATGGCAGCCCCGTCAAGGCGGG AGTGGAGACCACCAAACCCTCCAAACAGA GCAACAACAAGTACGCGGCCAGCAGCTACC TGAGCCTGACGCCCGAGCAGTGGAAGTCCC ACAGAAGCTACAGGTGCCAGGTCACGCATG AAGGGAGCACCGTGGAGAAGAGAGTGGCC CCTACAGAATGTTCA 410

318

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: Jail CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCrTC TCI GGGTTCTC ACTCAACA ATGCTAGAATGGGTGTGAGCTGGATCCGTC AGCCCCCAGGGAAGGCCCTGGAGTGGCTTG CACACATTTTTTCGAATGACGAAAAATCCT ACAGCACATCTCTGAAGAGCAGGCTCACCA TCTCCAAGGACACCTCCAAAAGCCAGGTGG TCCTAATTATGACCAACATGGACCCTGTGG ACACAGCCACATATTACTGTGCACGGTCAG TAGTAACTGGCGGCTACTACTACGACGGTA TGGACGTCTGGGGCCAAGGGACCAÇGGTCA CCGTCTCTAGTGCCTCCACCAAGGGCCCAT CGGTCTTCCCCCTGGCGCCCTGCTCCAGGA GCACCTCCGAGAGCACAGCGGCCCTGGGCT GCCTGGTCAAGGACTACTTCCCCGAACCGG TGACGGTGTCGTGGAACTCAGGCGCTCTGA CCAGCGGCGTGCACACCTTCCCAGCTGTCC TACAGTCCTCAGGACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCG GCACCCAGACCTACACCTGCAACGTAGATC ACAAGCCGAGCAACACCAAGGTGGACAAG ACAGTTGAGCGCAAATGTTGTGTCGAGTGC CCACCGTGCCCAGCACCACCTGTGGCAGGA CCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTG AGGTCACGTGCGTGGTGGTGGACGTGAGCC ACGAAGACCCCGAGGTCCAGTTCAACTGGT ACGTGGACGGCGTGGAGGTGCATAATGCCA AGACAAAGCCACGGGAGGAGCAGTTCAAC heavy from 16H7 V24F AGCACGTTCCGTGTGGTCAGCGTCCTCACC GTTGTGCACCAGGACTGGCTGAACGGCAAG GAGTACAAGTGCAAGGTCTCCAACAAAGGC CTCCCAGCCCCCATCGAGAAAACCATCTCC AAAACCAAAGGGCAGCCCCGAGAACCACA GGTGTACACCCTGCCCCCATCCCGGGAGGA GATGACCAAGAACCAGGTCAGCCTGACCTG CCTGGTCAAAGGCTTCTACCCCAGCGACAT CGCCGTGGAGTGGGAGAGCAATGGGCAGC CGGAGAACAACTACAAGACCACACCTCCCA TGCTGGACTCCGACGGCTCCTTCTTCCTCTA CAGCAAGCTCACCGTGGACAAGAGCAGGT GGCAGCAGGGGAACGTCTTCTCATGCTCCG TGATGCATGAGGCTCTGCACAACCACTACA CGCAGAAGAGCCTCTCCCTGTCTCCGGGTA AA 411

319

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 heavy chain I83T CAGGTCACCTTGAAGGAGTCTGGTCCTGTG ctggtgaaacccacagagaccctcacgctG ACCIOCACXGTGTCTGGGTrCTCACTCAAC aatgctagaatgggtgtgagctggatccgt CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT gcacacattttttcgaatgacgaaaaatcc tacagcàcatctctgaagagcaggctcacc atctccaaggacacctccaaaagccaggtg gtcctaaccatgacca ac atggaccctgtg gacacagccacatattactgtgcacggtca gtagtaactggcggctactactacgacggt atggacgtctggggccaagggaccacggtc accgtctctagtgcctccaccaagggccca tcggtcttccccctggcgccctgctccagga gcacctccgagagcacagcggccctgggct gcctggtcaaggactacttccccgaaccgg tgacggtgtcgtggaactcaggcgctctga CCAGCGGCGTGCACACCTTCCCAGCTGTCC TACAGTCCTCAGGACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCG GCACCCAGACCTACACCTGCAACGTAGATC ACAAGCCCAGCAACACCAAGGTGGACAAG ACAGTTGAGCGCAAATGTTGTGTCGAGTGC CCACCGTGCCCAGCACCACCTGTGGCAGGA CCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTG AGGTCACGTGCGTGGTGGTGGACGTGAGCC ACGAAGACCCCGAGGTCCAGTTCAACTGGT ACGTGGACGGCGTGGAGGTGCATAATGCCA AGACAAAGCCACGGGAGGAGCAGTTCAAC AGCACGTTCCGTGTGGTCAGCGTCCTCACC GTTGTGCACCAGGACTGGCTGAACGGCAAG gagta caagtgcaaggtctccaacaaaggc CTCCCAGCCCCCATCGAGAAAACCATCTCC aaaaccaaagggcagccccgagaaccaca ggtgtacaccctocccccatcccgggagga gatgaccaagaaccaggtcagcctgacctg cctggtcaaaggcttctaccccagcgacat cgccgtggagtgggagagcaatgggcagc CGGAGAACAACTACAAGACCACACCTCCCA TGCTGGACrCCGACGGCTCCTTCrrCCTCTA CAGCAAGCTCACCGTGGACAAGAGCAGGT ggcagcaggggaaçgtcttctcatgctccg TGATGCATGAGGCTCTGCACAACCACTACA cgcagaagagcctctccctgtctccgggta AA. _; 412

320

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 heavy chain V24F + I83T CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCTTÇTCTGGGTTCTCACTCAACA ATGCTAGAATGGGTGTGAGCTGGATCCGTC AGCCCCCAGGGAAGGCCCTGGAGTGGCTTG CACACATTTTTTCGAATGACGAAAAATCCT ACAGCACATCTCTGAAGAGCAGGCTCACCA tctccaaggacacctccaaaagccaggtgg TCCTAACCATGACCAACATGGACCCTGTGG ACACAGCCACATATTACTGTGCACGGTCAG TAGTAACTGGCGGCTACTACTACGACGGTA TGGACGTCTGGGGCCAAGGGACCACGGTCA CCGTCTCTAGTGCCTCCACCAAGGGCCCAT CGGTCTTCCCCCTGGCGCCCTGCTCCAGGA GCACCTCCGAGAGCACAGCGGCCCTGGGCT GCCTGGTCAAGGACTACTTCCCCGAACCGG TGACGGTGTCGTGGAACTCAGGCGCTCTGA CCAGCGGCGTGCACACCTTCCCAGCTGTCC TACAGTCCTCAGGACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCG GCACCCAGACCTACACCTGCAACGTAGATC ACAAGCCCAGCAACACCAAGGTGGACAAG ACAGTTGAGCGCAAATGTTGTGTCGAGTGC CCACCGTGCCCAGCACCACCTGTGGCAGGA CCGTCAGTCTTCCTCTTGCCGCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTG AGGTCACGTGCGTGGTGGTGGACGTGAGCC ACGAAGACCCCGAGGTCCAGTTCAACTGGT ACGTGGACGGCGTGGAGGTGCATAATGCCA AGACAAAGCCACGGGAGGAGCAGTTCAAC AGCACGTTCCGTGTGGTCAGCGTCCTCACC GTTGTGCACCAGGACTGGCTGAACGGCAAG GAGl'ACAAGTGCAAGGTCl'CCAACAAAGGC CTCCCAGCCCCCATCGAGAAAACCATCTCC AAAACCAAAGGGCAGCCCCGAGAAÇCACA GGTGTACACCCTGCCCCCATCCCGGGAGGA GATGACCAAGAACCAGGTCAGCCTGACCTG CCTGGTCAAAGGCTTCTACCCCAGCGACAT CGCCGTGGAGTGGGAGAGCAATGGGCAGC CGGAGAACAACTACAAGACCACACCTCCCA TGCTGGACTCCGACGGCTCCTTCTTCCTCTA CAGCAAGCTCACCGTGGACAAGAGCAGGT GGCAGCAGGGGAACGTCTTCTCATGCTCCG TGATGCATGAGGCTCTGCACAACCACTACA cgcagaagagcctctccctgtctccgggta AA 413

321

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 heavy chain E16Q + V24F + I83T CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAÇAGACCCTCACGCTG ACCTGCACCTTCTCTGGGTTCTCACTCAACA ÀTGCTAGAATGGGTGTGAGCTGGATCCGTC AGCCCCCAGGGAAGGCCCTGGAGTGGCTTG CÀCáCAITTTTTCGAATGACGAAAAATCCT ACAGCAC'ATCTCTGAAGAGCAGGCTCACCA TCTCCAAGGACACCTCCAAAAGCCAGGTGG TCCTAACCATGACCAACATGGACCCTGTGG ACACAGCCACATAITACTGTGCACGGTÇAG TAGTAACTGGCGGCTACTACTACGACGGTA TGGACOTCTGGGGCCAAGGGACCAÇGGTCA CCGTCTCTAGTGCCTCCACCAAGGGCCCAT CGGTCTTCCCCCTGGCGCCCTGCTCCAGGA GCACCTCCGAGAGCACAGCGGCCCTGGGCT GCCTGGTCAAGGACTACTTCCCCGAACCGG TGACGGTGTCGTGGAACTCAGGCGCTCTGA CCAGCGGCGTGCACACCTTCCCAGCTGTCC TACAGTCCTCAGGACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCG GCACCCAGACCTACACCTGCAACGTAGATC ACAAGCCCAGCAACACCAAGGTGGACAAG acagttgagcgcaaatgttgtgtcgagtgc CCACCGTGCCCAGCACCACCTGTGGCAGGA CCGTCAGTCTTCCTCTrCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTG AGGTeACGTGCGTGGTGGTGGACGTGAGCC ACGAAGACCCCGAGGTCCAGTTCAACTGGT ACGTGGACGGCGTGGAGGTGCATAATGCCA AGACAAAGCCACGGGAGGAGCAGTTCAAC AGCAGGTTCCGTGTGGTCAGCGTCCTCACC GTTGTGCACCAGGACTGGCTGAACGGCAAG L AGTACAAGTGCAAGGTCTCCAACAAAGGC CTCCCAGCCCCCATCGAGAAAACCATCTCC AAAACCAAAGGGCAGCCCCGAGAACCACA GGTGTACACCCTGCCCCCATCCCGGGAGGA GATGACCAAGAACCAGGTCAGCCTGACCTG CCTGGTCAAAGGCTTCTACCCCAGCGACAT CGCCGTGGAGTGGGAGAGCAATGGGCAGC CGGAGAACAACTACAAGACCACACCTCCCA TGCTGGACTCCGACGGCTCCTTCTTCCTCTA CAGCAAGCTCACCGTGGACAAGAGCAGGT GGCAGCAGGGGAACGTCTTCTCATGCTCCG TGATGCATGAGGCTCTGCACAACCACTACA CGCAGAAGAGCCTCTCCCTGTCTCCGGGTA AA 414

322

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 heavy chain E16Q + V24F + I83T + T119L CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAÇAGACCCTCACGCTG ACCTGCACCTTCTCTGGGTTCTCACTCAACA ATGCTAGAATGGGTGTGAGCTGGATCCGTC AGCCCCCAGGGAAGGCCCTGGAGTGGCTTG C ACGA ACAC ATTTTTTCGAATG AAAATCCT ACAGCACATCTCTGAAGAGCAGGCTCACCA TCTCCAAGGACACCTCCAAAAGCCAGGTGG TCCTAACCATGACCAACATGGACCCTGTGG ACACAGCCACATATTACTGTGCACGGTCAG tagtaactggcggctactactacgacggta TGGACGTCTGGGGCCAAGGGACCÇTCGTCA ccgtctctagtgcctccaccaagggcccat cggtcttccccctggcgccctgctccagga gcacctccgagagcacagcggccctgggct gcctggtcaaggactacttccccgaaccgg tgacggtgtcgtggaactcaggcgctctga ccagcggcgtgcacaccttcccágctgtcc tacagtcctcaggactctactccctcagca gcgtggtgaccgtgccctccagcaacttcg gcacccagacctacacctgcaacgtagatc acaagcccagcaacaccaaggtggacaag acagttgagcgcaaatgttgtgtcgagtgc ccaccgtgcccagcaccacctgtggcagga ccgtcagtcttcctcttccccccaaaaccca aggacaccctcatgatctcccggacccctg aggtcacgtgcgtggtggtggacgtgagcc acgaagaccccgaggtccagttcaactggt acgtggacggcgtggaggtgcataatgcca agacaaagccacgggaggagcagttcaac agcacgttccgtgtggtcagcgtcctcacc gttgtgcagcaggactggctgaacggcaag L AGTACAAGTGCAAGGTCTCCAACAAAGGC CTCCCAGCGCCCATCGAGAAAACCATCTCC aaaaccaaagggcagccccgagaaccaca GGTGTACACCCTGCCCCCATCCCGGGAGGA GATGACCAAGAACCAGGTCAGCCTGACCTG CCTGGTCAAAGGCTTCTACCCCAGCGACAT cgccgtggagtgggagagcaatgggcagc cggagaacaactacaagaccacacctccca TGCTGGACTCCGACGGCTCCnTCTTCCTCTA CAGCAAGCTCACCGTGGACAAGAGCAGGT GGCAGCAGGGGAACGTCTTCTCATGCTCCG TGATGCATGAGGCTCTGCACAACCACTACA CGCAGAAGAGCCTCTCCCTGTCTCCGGGTA A A 415

323

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 heavy chain E16Q + V24F + I83T + S100I + T119L CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACACAGACCCTCACGCTG ACCTGCACCTTCTCTGGGTTCTCACTCAACA ATGCTAGAATGGGTGTGAGCTGGATCCGTC AGCCCCGAGGGAAGGCCCTGGAGTGGCTTG CACACATTTTTTCGAATGACGAAAAATCCT ACAGCACATCTCTGAAGAGCAGGCTCACCA TCTCCAAGGACACCTCCAAAAGCCAGGTGG TCCTAACCATGACCAACATGGACCCTGTGG ACACAGCCACATATTACTGTGCACGGATCG TAGTAACTGGCGGCTACTACTACGACGGTA TGGACGTCTGGGGCCAAGGGACCCTGGTCA CCGTCTCTAGTGCCTCCACCAAGGGCCCAT CGGTCTTCCCCCTGGCGCCCTGCTCCAGGA GCACCTCCGAGAGCACAGCGGCCCTGGGCT GCCTGGTCAAGGACTACTTCCCCGAACCGG TGACGGTGTCGTGGAACTCAGGCGCTCTGA CCAGCGGCGTGCACACCTTCCCAGCTGTCC TACAGTCCTCAGGACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCG GCACCCAGACCTACACCTGCAACGTAGATC ACAAGCCCAGCAACACCAAGGTGGACAAG ACAGTTGAGCGCAAATGTTGTGTCGAGTGC CCACCGTGCCCAGCACCACCTGTGGCAGGA CCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTG AGGTCACGTGCGTGGTGGTGG ACGTGAGCC ACGAAGACCCCGAGGTCCAGTTCAACTGGT ACGTGGACGGCGTGGAGGTGCATAATGCCA AGACAAAGCCACGGGAGGAGCAGTTCAAC AGCACGTTCCGTGTGGTCAGCGTCCTCACC GTrGTGCAGCAGGACTGGCTGAACGGCAAG GAGTACA AGTGCAAGGTCTCCAACAAAGGC CTCCCAGCCCCCATCGAGAAAACCATCTCC AAAACCAAAGGGCAGCCCCGAGAACCACA GGTGTACACCCrGCCCCCATCCCGGGAGGA GATGACCAAGAACCAGGTCAGCCTGACCTG CCTGGTCAAAGGCTTCTACCCCAGCGACAT CGCCGTGGAGTGGGAGAGCAATGGGCAGC CGGAGAACAACTACAAGACCACACCTCCCA TGCTGGACl'CCGACGGCTCCn'CTTCCTCTA CAGCAAGCTCACCGTGGACAAGAGCAGGT GGCAGCAGGGGAACGTCTTCTCATGCTCCG TGATGCATGAGGCTCTGCACAACCACTACA CGCAGAAGAGCCTCTCCCTGTCTCCGGGTA AA 416

324

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 heavy chain I83K CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCGTGTCTGGGTTCTCACTCAAC AATGCTAGAATGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACGAAAAATCC TACAGCACATCTCTGAAGAGCAGGCTCACC ATCTCCAAGGACACCTCCAAAAGCCAGGTG GTCCTAAAGATGACCAACATGGACCCTGTG GACACAGCCACATATTACTGTGCACGGTCA GTAGTAACTGGCGGCTACTACTACGACGGT ATGGACGTCTGGGGCCAAGGGACCACfiGTC ACCGTCTCTAGTGCCTCCACCAAGGGCCCA TCGGTCTTCCCCCTGGCGCCCTGCTCCAGGA GCACCTCCGAGAGCACAGCGGCCCTGGGCT GCCTGGTCAAGGACTACTTCCCCGAACCGG TGACGGTGTCGTGGAACTCAGGCGCTCTGA CCAGCGGCGTGCACACCTTCCCAGCTGTCC TACAGTCCTCAGGACl'CTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCG GCACCCAGACCTACACCTGCAACGTAGATC ACAAGCCCAGCAACACCAAGGTGGACAAG ACAGTTGAGCGCAAATGTTGTGTCGAGTGC CCAÇCGTGCCCAGCACCACCTGTGGCAGGA CCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTG AGGTCACGTGCGTGGTGGTGGACGTGAGCC ACGAAGACCCCGAGGTCCAGTTCAACTGGT ACGTGGACGGCGTGGAGGTGCATAATGCCA AGACAAAGCCACGGGAGGAGCAGTTCAAC AGCACGTTCCGTGTGGTCAGCGTCCTCACC GTTGTGCACCAGGACTGGCTGAACGGCAAG GAGTACAAGTGCAAGGTCTCCAACAAAGGC CTCCCAGCCCCCATCGAGAAAACCATCTCC AAAACCAAAGGGCAGCCCCGAGAACCACA GGTG TACACCCTGCCCCCATCCCGGGAGGA GATGACCAAGAACCAGGTCAGCCTGACCTG CCTGGTCAAAGGCrrCTACCCCAGCGACAT CGÇCGTGGAGTGGGAGAGCAATGGGCAGC CGGAGAACAACTACAAGACCACACCTCCCA TGCTGGACTCCGACGGCTCCTTCTTCCTCTA CAGCAAGCTCACCGTGGACAAGAGCAGGT GGCAGCAGGGGAACGTCTTCTCATGCTCCG TGATGCATGAGGCTCTGCACAACCACTACA CGCAGAAGAGCCrCTCCCTGTCTCCGGGTA AA 417

325

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 heavy chain S100I CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGÀCCCTCACGCTG ACCTGCACCGTGTCTGGGTTCTCACTCAAC AATGCTAGAATGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACGAAAAATCC TACAGCACATCTCTGAAGAGCAGGCTCACC ATCTCCAAGGACACCrCCAAAAGCCAGGTG GTCCT AATT ATG ACC AAC ATGG ACCCTGTG GACACAGCCACATATTACTGTGCACGGATC GTAGTAACTGGCGGCTACTACTACGACGGT ATGGACGTCTGGGGCCAAGGGACCACGGTC ACCGTCTCTAGTGCCTCCACCAAGGGCCCA TCGGTCTTCCCCCTGGCGCCCTGCTCCAGGA GCACCTCCGAGAGCACAGCGGCCCTGGGCT GCCTGGTCAAGGACTACTTCCCCGAACCGG TGACGGTGTCGTGGAACTCAGGCGCTCTGA CCAGCGGCGTGCACACCTTCCCAGCTGTCC TACAGTCCTCAGGACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCG GCACCCAGACCTACACCTGCAACGTAGATC ACAAGCCCAGCAACACCAAGGTGGACAAG ACAGTTGAGCGCAAATGTTGTGTCGAGTGC CCACCGTGCCCAGCACCACCTGTGGCAGGA CCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTG AGGTCACGTGCGTGGTGGTGGACGTGAGCC ACGAAGACCCCGAGGTCCAGTTCAACTGGT ACGTGGACGGCGTGGAGGTGCATAATGCCA AGACAAAGCCACGGGAGGAGCAGTTCAAC AGCACGTTCCGTGTGGTCAGCGTCCTCACC GTTGTGCACCAGGACTGGCTGAACGGCAAG L AGTACAAGTGCAAGGTCTCCAACAAAGGC CTCCCAGCCCCCATCGAGAAAACCATCTCC AAAACCAAAGGGCAGCCCCGAGAACCACA GGTGTACACCCTGCCCCCATCCCGGGAGGA GATGACCAAGAACCAGGTCAGÇCTGACCTG cctggtcaaaggcttctaccccagcgacat CGCCGTGGAGTGGGAGAGCAATGGGCAGC cggagaacaactacaagaccacacctccca tgctggactccgacggctccttcttcctcta cagcaagctcaccgtggacaagagcaggt ggçagcaggggaacgtcttctcatgctccg tgatgcatgaggctctgcacaaccactaca CGCAGAAGAGCCrCTCCCTGTCTCCGGGTA ΑΑ 418

326

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 heavy chain D88R + P8 9A + V90E CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCGTGTCTGGGTTCrCACTCAAC AATGCTAGAATGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACGAAAAATCC TACAGCACATCTCTGAAGAGCAGGCTCACC ATCTCCAAGGACACCTCCAAAAGCCAGGTG GTCCTAATTATGACCAACATGAGAGCTGAG GACACAGCCACATATTACTGTGCACGGTCA GTAGTAACTGGCGGCTACTACTACGACGGT ATGGACGTCTGGGGCCAAGGGACCACGGTC ACCGTCTCTAGTGCCTCCACCAAGGGCCCA TCGGTCTTCCCCCTGGCGCCCTGCTCCAGGA GCACCTCCGAGAGCACAGCGGCCCTGGGCT GCCTGGTCAAGGACTACTTCCCCGAACCGG TGACGGTGTCGTGGAACTCAGGCGCTCTGA CCAGCGGCGTGCACACCTTCCCAGCTGTCC TACAGTCCTCAGOACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCG GCACCCAGACCTACACCTGCAACGTAGATC ACAAGCCCAGCAACACCAAGGTGGACAAG ACAGTTGAGCGCAAATGTTGTGTCGAGTGC CCACCGTGCCCAGCACCACCTGTGGCAGGA CCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTG AGGTCACGTGCGTGGTGGTGGACGTGAGCC ACGAAGACCCCGAGGTCCAGTTCAACTGGT ACGTGGACGGCGTGGAGGTGCATAATGC'CA AGACAAAGCCACGGGAGGAGCAGTTCAAC AGCACGTTCCGTGTGGTCAGCGTCCTCACC GTTGTGCACCAGGACTGGCTGAACGGCAAG GAGTACA AGTGCAAGGTCTCCAACAAAGGC CTCCCAGCCCCCATCGAGAAAACCATCTCC AAAACCAAAGGGCAGCCCCGAGAACCACA GGTGTACACCCTGCCCCCATCCCGGGAGGA GATGACCAAGAACCAGGTCAGCCTGACCTG CCTGGTCAAAGGCTTCTACCCCAGCGACAT CGCCGTGGAGTGGGAGAGCAATGGGCAGC CGGAGAACAACTACAAGACCÀCACCTCCCA TGCTGGACTCCGACGGCTCCTTCTTCCTCTA CAGCAAGCTCACCGTGGACAAGAGCAGGT GGCAGCAGGGGAACGTCTTCTCATGCTCCG TGATGCATGAGGCTCTGCACAACCACTACA CGCAGAAGAGCCTCTCCCTGTCTCCGGGTA AA 419

327

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 heavy chain D88R + P8 9A + V90E + S100I CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAOAGACCCTCACGCTG ACCTGCACCGTGTCTGGGTTCTCACTCAAC AATGCTAGAATGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACGAAAAATCC TACAGCACATCTCTGAAGAGCAGGCTCACC ATCTCCAAGGACACCTCCAAAAGCCAGGTG GTCCTAATTATGACCAACATGAGAGCTGAG GACACAGCCACATATTACTGTGCACGGATÇ GTAGTAACTGGCGGCTACTACTACGACGGT ATGGACGTCTGGGGCCAAGGGACCACGGTC ACCGTCTCTAGTGCCTCCACCAAGGGCCCA TCGGTCTTCCCCCTGGCGCCCTGCTCCAGGA GCACCTCCGAGAGCACAGCGGCCCTGGGCT GCCTGGTCAAGGACTACTTCCCCGAACCGG TGACGGTGTCGTGGAACTCAGGCGCTCTGA CCAGCGGCGTGCACACCTTCCCAGCTGTCC TACAGTCCTCAGGACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCG GCACCCAGACCTACACCTGCAACGTAGATC ACAAGCCCAGCAACACCAAGGTGGACAAG ACAGTTGAGCGCAAATGTTGTGTCGAGTGC CCAÇCGTGCCCAGCACCACCTGTGGCAGGA CCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTG AGGTCACGTGCGTGGTGGTGGACGTGAGCC ACGAAGACCCCGAGGTCCAGTTCAACTGGT ACGTGGACGGCGTGGAGGTGCATAATGCCA agacaaagccacgggaggagcagttcaac AGCACGITCCGTGTGGTCAGCGTCCTCACC GTTGTGCACCAGGACTGGCTGAACGGCAAG GAGTAC AAGTGCAAGGTCTCCAACAAAGGC CTCCCAGCCCCCATCGAGAA ACC ATCTCC AAAÁCCAAAGGGCAGCCCCGAGAACCACA GGTGTACACCCTGCCCCCATCCCGGGAGGA GATGACCAAGAACCAGGTCAGCCTGACCTG CCTGGTCAAAGGCTTCTACCCCAGCGACAT CGCCGTGGAGTGGGAGAGCAATGGGCAGC CGGAGAACAACTACAAGACCACACCTCCCA TGCTGGACTCCGACGGCTCCTTCTTCCTCTA CAGCAAGCTCACCGTGGACAAGAGCAGGT GGCAGCAGGGGAACGTCTTCTCATGCTCCG TGATGCATGAGGCTCTGCACAACCACTACA CGCAGAAGAGCCTCTCCCTGTCTCCGGGTA AA 420

328

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 heavy chain Y107 deletion CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCGTGTCTGGGTTCTCACTCAAC aatgctagaatgggtgtgagctggatccgt CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT AC AC GC GC AATGACG The HIM AAAATCC TACAGCACATCTCTGAAGAGCAGGCTCACC ATCTCCAAGGACACCTCCAAAAGCCAGGTG GTCCIAATTATGACCAACATGGACCCTGTG GAÇACAGCCACATATTACTGTGCACGGTÇA GTAGTAACTGGCGGCTACITACGACGGTATG GACGTCTGGGGCCAAGGGACCAÇGGTCACC GTCTCTAGTGCCTCCACCAAGGGCCCATCG GTCTTCCCCCTGGCGCCCTGCTCCAGGAGC ACCTCCGAGAGCACAGCGGCCCTGGGCTGC CTGGTCAAGGACTACTTCCCCGAACCGGTG ACGGTGTCGTGGAACTCAGGCGCTCTGACC AGCGGCGTGCACACCTTCCCAGCTGTCCTA CAGTCCTCAGGACTCTACTCCCTCAGCAGC GTGGTGACCGTGCCCTCCAGCAACTTCGGC ACCCAGACCTACACCTGCAACGTAGATCAC AAGCCCAGCAACACCAAGGTGGACAAGAC AGTTGAGCGCAAATGTTGTGTCGAGTGCCC ACCGTGCCCAGCACCACCTGTGGCAGGACC GTCAGTCTTCCTCTTCCCCCCAAAACCCAAG GACACCCTCATGATCTCCCGGACCCCTGAG GTCACGTGCGTGGTGGTGGACGTGAGCCAC GAAGACCCCGAGGTCCAGTTCAACTGGTAC GTGGACGGCGTGGAGGTGCATAATGCCAAG ACAAAGCCACGGGAGGAGCAGTTCAACAG CACGTTCCGTGTGGTCAGCGTCCTCACCGTT GTGCACCAGGACTGGCTGAACGGCAAGGA GTACAAGTGCAAGGTCTCCAACAAAGGCCT CCCAGCCCCCATCGAGAAAACCATCTCCAA AACCAAAGGGCAGCCCCGAGAACCACAGG TGTACACCCTGCCCCCATCCCGGGAGGAGA TGACCAAGAACCAGGTCAGCCTGACCTGCC TGGTCAAAGGCTrCTACCCCAGCGACATCG CCGTGGAGTGGGAGAGCAATGGGCAGCCG GAGAACAACTACAAGACCACACCTCCCATG CTGGACTCCGACGGCTCCTTCTTCCTCTACA GCAAGCTCACCGTGGACAAGAGCAGGTGG CAGCAGGGGAACGTCTrCTCATGCTCCGTG ATGCATGAGGCTCTGCACAACCACTACACG CAGAAGAGCCTCTCCCTGTCTCCGGGTAAA 421

329

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 16H7 heavy chain D109S CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCGTGTCTGGGTTCTCACTCAAC AATGCTAGAATGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACGAAAAATCC TACAGCACATCTCTGAAGAGCAGGCTCACC ATCTCCAAGGACACCTCCAAAAGCCAGGTG GTCCTAATTATGACCAACATGGACCCTGTG GACACAGCCACATATTACTGTGCACGGTCA GTAGTAACTGGCGGCTACTACTACAGCGGT ATGGACGTCTGGGGCCAAGGGACCAÇGGTC ACCGTCTCTAGTGCCTCCACCAAGGGCCCA TCGGTCTTCCCCCTGGCGCCCTGCTCCAGGA GCACCTCCGAGAGCACAGCGGCCCTGGGCT GCCTGGTCAAGGACTACTTCCCCGAACCGG TGACGGTGTCGTGGAACTCAGGCGCTCTGA CCAGCGGCGTGCACACCTTCCCAGCTGTCC TACAGTCCTCAGGACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCG GCACCCAGACCTACACCTGCAACGTAGATC ACAAGCCCAGCAACACCAAGGTGGACAAG ACAGTTGAGCGCAAATGTTGTGTCGAGTGC CCACCGTGCCCAGCACCACCTGTGGCAGGA CCGTCAGTCTTCCTCTTCCCCCCAAAACCCA aggacaCcctcatgatctcccggacccctg aggtcaCgtgcgtggtggtggacgtgagcc acgaagaccccgaggtccagttcaactggt acgtggacggcgtggaggtgcataatgcca agacaaagccacgggaggagcagttcaac agcacgttccgtgtggtcagcgtcctcacc gttgtgcaccaggactggctgaacggcaag oagtaca agtgcaaggtctccaacaaaggc ctcccagcccccatcgagaaaaccatctcc aaaaccaaagggcagccccgagaaccaca ggtgtacaccctgcccccatcccgggagga gatgaccaagaaccaggtcagcctgacctg Cctggtcaaaggcttctaccccagcgacat cgccgtggagtgggagagcaatgggcagc cggagaacaactacaagaccacacctccca tgctggactccgacggctccttcttcctcta cagcaagctcaccgtggacaagagcaggt ggcagcaggggaacgtcttctcatgçtccg tgatgcatgaggctctgcacaaccactaca cgcagaagagcctctccctgtçtccgggta AA 422

330

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: Jail take of 22H5 N92Q TCCTATGTGCTGACTCAGCCACCCTCGGTGT CAGTGGCCCCAGGACAGACGGCCAGGATTA cctgtgggggaaacaacattggaagtcaaa GTGTGCACTGGTACCAGCAGAAGCCAGGCC AGGCCCCTGTCCTGGTCGTCTATGATGATA GCGACCGGCCCTCAGGGATCCCTGAGCGAT TCTCTGGTTCCAACTCTGGGAACACGGCCA CCCTGACCATCAGCAGGGTCGAAGCCGGGG ATGAGGCCGACTATTACTGTCAGGTGTGGG ATCAGACTAGTGATCATGTGGTATTCGGCG GGGGGACCAAGCTGACCGTCCTAGGTCAGC CCAAGGCCAACCCCACTGTCACTCTGTTCC CGCCCTCCTCTGAGGAGCTCCAAGCCAACA AGGCCACACTAGTGTGTCTGATCAGTGACT TCTACCCGGGAGCTGTGACAGTGGCCTGGA AGGCAGATGGCAGCCCCGTCAAGGCGGGA GTGGAGACCACCAAACCCTCCAAACAGAGC AACAACAAGTACGCGGCCAGCAGCTACCTG AGCCTGACGCCCGAGCAGTGGAAGTCCCAC AGAAGCTACAGCTGCCAGGTCACGCATGAA GGGAGCACCGTGGAGAAGACAGTGGCCCC TACAGAATGTTCA 423 22H5 light chain S94A TCCTATGTGCTGACTCAGCCACCCTCGGTGT CAGTGGCCCCAGGACAGACGGCCAGGATTA CCTGTGGGGGAAACAACATTGGAAGTCAAA GTGTGCACTGGTACCAGCAGAAGCCAGGCC AGGCCCCTGTCCTGGTCGTCTATGATGATA GCGACCGGCCCTCAGGGATCCCTGAGCGAT TCTCTGGTTCCAACTCTGGGAACACGGCCA CCCTGACCATCAGCAGGGTCGAAGCCGGGG ATGAGGCCGACTATTACTGTCAGGTGTGGG ATAATACTGCTGATCATGTGGTAITCGGCG GGGGGACCAAGCTGACCGTCCTAGGTCAGC CCAAGGCCAACCCCACTGTCACTCTGTTCC CGCCCTCCTCTGAGGAGCTCCAAGCCAACA AGGCCACACTAGTGTGTCTGATCAGTGACT TCTACCCGGGAGCTGTGACAGTGGCCTGGA AGGCAGATGGCAGCCCCGTCAAGGCGGGA GTGGAGACCACCAAACCCTCCAAACAGAGC AACAACAAGTACGCGGCCAGCAGCTACCTG AGCCTGACGCCCGAGCAGTGGAAGTCCCAC AGAAGCTACAGCTGCCAGGTCACGCATGAA GGGAGCACCGTGGAGAAGACAGTGGCCCC lACAGAATGriCA 424

331

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 22H5 heavy chain C109S CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCGTGTCTGGGTTCTCACTCAGC AATGCTAGAATGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACGAAAAATCC TACAGCACATCTCTGAAGAGCAGGCTCACC ATCTCCAAGGACACCTCCAAAAGCCAGGTG GTCCTTACCATGACCAACATGGACCCTGTG GACACAGCCACATATTACTGTGCACGGATA TTATTAGTGGGAGCTTACTACTACAGCGGT ATGGACGTCTGGGGCCAAGGGACCACGGTC ACCGTCTCTAGTGCCTCCACCAAGGGCCCA TCGGTCTTCCCCCTGGCGCCCTGCTCCAGGA GCÁCCTCCGAGAGCACAGCGGCCCTGGGCT GCCTGGTCAAGGACTACTTCCCCGAACCGG TGACGGTGTCGTGGAÂCTCAGGCGCTCTGA CCAGCGGCGTGCACACCTTCCCAGCTGTCC TACAGTCCTCAGGACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCG GCACCCAGACCTACACCTGCAACGTAGATC ACAAGCCCAGCAACACCAAGGTGGACAAG ACAGTTGAGCGCAAATGTTGTGTCGAGTGC CCACCGTGCCCAGCACCACCTGTGGCAGGA CCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTG AGGTCACGTGCGTGGTGGTGGACGTGAGCC ACGAAGACCCCGAGGTCCAGTTCAACTGGT ACGTGGACGGCGTGGAGGTGCATAATGCCA AGACAAAGCÇACGGGAGGAGCAGTTCAAC AGCACGTTCCGTGTGGTCAGCGTCCTCACC GTTGTGCACCAGGACTGGCTGAACGGCAAG GAGTA CAAGTGCAAGGTCTCCAACAAAGGC CTCCCAGCCCCCATCGAGAAAACCATCTCC AAAACCAAAGGGCAGCCCCGAGAACCACA GGTGTACACCCTGCCCCCATCCCGGGAGGA GATGACCAAGAACCAGGTCAGCCTGACCTG CCTGGTCAAAGGCTTCTACCCCAGCGACAT CGCCGTGGAGTGGGAGAGCAATGGGCAGC CGGAGAACAACTACAAGACCACACCTCCCA TGCTGGACTCCGACGGCTCCTTCTTCCTCTA CAGCAAGCTCACCGTGGACAAGAGCAGGT GGCAGCAGGGGAACGTCTTCTCATGCTCCG TGATGCATGAGGCTCTGCACAACCACTACA CGCAGAAGAGCCTCTCCCTGTCTCCGGGTA AA. 425

332

Seq. central Variation Nucleic acid sequence ID. IN SEQ. N °: 22H5 heavy chain Y107 insertion CAGGTCACCTTGAAGGAGTCTGGTCCTGTG CTGGTGAAACCCACAGAGACCCTCACGCTG ACCTGCACCGTGTCTGGGTTCTCACTCAAC AATGCTAGAATGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCTGGAGTGGCTT GCACACATTTTTTCGAATGACGAAAAATCC TACAGCACATCTCTGAAGAGCAGGCTCACC ATCTCCAAGGACACCTCCAAAAGCCÀGGTG GTCCTAATTATGACCAACATGGACCCTGTG GACACAGCCACATATTACTGTGCACGGTÇA GTAOTAACTGGCGGCTACTATTACTACGAC GGTATGGACGTCTGGGGCCAAGGGACCACG GTCACCGTCTCTAGTGCCTCCACCAAGGGC CCATCGGTCTTCCCCCTGGCGCCCTGCTCCA GGAGCACCTCCGAGAGCACAGCGGCCCTGG GCTGCCTGGTCAAGGACTACTTCCCCGAAC CGGTGACGGTGTCGTGGAACTCAGGCGCTC TGACCAGCGGCGTGCACACCTTCCCAGCTG TCCTACAGTCCTCAGGACTCTACTCCCTCAG CAGCGTGGTGACCGTGCCCTCCAGCAACTT CGGCACCCAGACCTACACCTGCAACGTAGA TCACAAGCCCAGCAACACCAAGGTGGACA AGACAGTTGAGCGCAAATGTTGTGTCGAGT GCCCACCGTGCCCAGCACCACCTGTGGCAG GACCGTCAGTCTTCCTCTTCCCCCCAAAACC CAAGGACACCCTCATGATCTCCCGGACCCC TGAGGTCACGTGCGTGGTGGTGGACGTGAG CCACGAAGACCCCGAGGTCCAGTTCAACTG GTACGTGGACGGCGTGGAGGTGCATAATGC CAAGACAAAGCCACGGGAGGAGCAGTTCA ACAGCACGTTCCGTGTGGTCAGCGTCCTCA CCGTTGTGCACCAGGACTGGCTGAACGGCA AGGAG TACAAGTGCAAGGTCTCCAACAAA GGCCTCCCAGCCCCCATCGAGAAAACCATC TCCAAAACCAAAGGGCAGCCCCGAGAACC ACAGGTGTACAGCCTGCCCCCATCCCGGGA GGAGATGACCAAGAACCAGGTCAGCCTGA CCTGCCTGGTCAAAGGCTTCTACCCCAGCG ACATCGCCGTGGAGTGGGAGAGCAATGGG CAGCCGGAGAACAACTACAAGACCACACCT CCCATGCTGGACTCCGACGGCTCCTTCTTCC TCTACAGCAAGCTCACCGTGGACAAGAGCA GGTGGCAGCAGGGGAACGTCTTCTCATGCT CCGTGATGCATGAGGCTCTGCACAACCACT ACACGCAGAAGAGCCTCTCCCTGTCTCCGG GTAAA 426

333

ID No. SEQ. DA CDR3 00 00 00 00 00 00 429 430 148 148 CDR3 i Μ Q co a ·> Si I o QVWDGNSDH W ÍU Q co Si 5 CX QVWAGNSDH W a Q co § £ r CX SWTGGYYY DGMDV SWTGGYYY DGMDV ID No. SEQ. DA CDR2 co 427 428 kO 427 POO 133 CDR2 DDSDRPS YDSDRPS ADSDRPS DDSDRPS ADSDRPS HIFSNDEKSY STSLKS HIFSNDEKSY STSLKS ID No. SEQ. DA CDR1 kO 166 166 166 kü kO 122 122 CDR1 GGNNIGSES VH GGNNIGSES VH co W co i ^g 0 0 GGNNIGSES VH GGNNIGSES VH NARMGVS NARMGVS Mutation location to Cu CDR2 CDR2 CDR3 CDR2, CDR3 The & the Cm Variation Q16K > H σ Q D49A D91A + . 3 3 rH σ> σ Q Q V24F I83T Center string 16H7 light chain 16H7 light chain 16H7 light chain 16H7 light chain 16H7 light chain 16H7 heavy chain 16H7 heavy chain

334

ID No. SEQ. DA CDR3 148 148 148 431 148 CDR3 SWTGGYYY DGMDV SWTGGYYY DGMDV SWTGGYYY DGMDV IWTGGYYY DGMDV SWTGGYYY ID No. SEQ. DA CDR2 133 133 133 133 133 CDR2 HIFSNDEKSY STSLKS HIFSNDEKSY STSLKS HIFSNDEKSY STSLKS HIFSNDEKSY STSLKS HIFSNDEKSY ID No. SEQ. DA CDR1 122 122 122 122 122 CDR1 NARMGVS NARMGVS NARMGVS NARMGVS NARMGVS Mutation location FW FW FW FW, CDR3 FW Variation V24F + I83T E16Q + V24F + I83T E16Q + V24F + I83T + T119L ^{+ + + +} Q OQ Η θ 2 co Fl H CN CO □ ^w > ^H ω ^H I83K Center string 16H7 heavy chain 16H7 heavy chain 16H7 heavy chain 16H7 heavy chain Heavy chain

335

ID No. SEQ. DA CDR3 432 148 433 434 435 436 CDR3 DGMDV IWTGGYYY DGMDV SWTGGYYY DGMDV IWTGGYYY DGMDV SWTGGYYD GMDV SWTGGYYY SGMDV QVWDQTSDH ID No. SEQ. DA CDR2 133 133 133 CO 133 kD CDR2 STSLKS HIFSNDEKSY STSLKS HIFSNDEKSY STSLKS HIFSNDEKSY STSLKS HIFSNDEKSY STSLKS HIFSNDEKSY STSLKS DDSDRPS ID No. SEQ. DA CDR1 122 122 122 122 122 <0 CDR1 NARMGVS NARMGVS NARMGVS NARMGVS NARMGVS GGNNIGSQS Mutation location CDR3 3 X FW, CDR3 CDR3 CDR3 CDR3 Variation S100I D88R + P89A + V90E D88R + P89A + V90E + S100I Y107 deletion D109S N92Q Center string from 16H7 16H7 heavy chain 16H7 heavy chain 16H7 heavy chain 16H7 heavy chain 16H7 heavy chain Light chain

336

ID No. SEQ. DA CDR3 437 438 439 CDR3 & QVWDNTADH W ILLVGAYYY SGMDV SWTGGYYY YDGMDV ID No. SEQ. DA CDR2 133 133 CDR2 DDSDRPS HIFSNDEKSY STSLKS HIFSNDEKSY STSLKS ID No. SEQ. DA CDR1 167 122 122 CDR1 _ ΗΛ GGNNIGSQS VH NARMGVS NARMGVS Mutation location CDR3 CDR3 CDR3 Variation cn ra C109S Y107 insertion Center string from 22H5 22H5 light chain 22H5 heavy chain 16H7 heavy chain

Q cn H CH ft M Q O O frog U kO Q 0 M O Q Q z CD Eh co ft Q 0 0 0 Eh 0 0 The Eh 0 0 Eh 0 Eh 0 H £ 0 0 Eh <2 kJ 0 0 Q . 0 H CH ft M Q O frog U Q 0 M O Q Q z CD 0 0 H 0 0 CN 0 ft Eh 0 Q CD 0 CD CD H CD < 0 0 Q • J — I H CH ft M Q cn O frog U CO Q 0 M Ç O Q Q z 0 H ft Q U GGGAAA 0 0 Eh Eh 0 0 1 H 0 0 0 Eh CD 0 kJ 0 0 ΙΦ O Λ N 0 i <0 U · <0 4J z P Φ 6 υ 0 J gives 0 1Φ O' ft <β kO rd j— | O' Φ > (U Φ > • rl | 0) 0 Φ p — j ft you P kD «D 4J rtS you • rd & 0) 0) (D 0) O T5 TS "1 Φ 0

337

d co Η ο Hi Μ Ω ο Q Ο ω Ο k0 ω CN CN W ο Ω Ω The ο d Εη 0 H H <η 0 Η 0 The < 0 Εη £ 0 Eh 0 H < 0 Eh <H Ρ5 Ε-ι 0 kJ Eh 0 kJ Q 0 0 0 kJ 0 CO CO kJ C-l 0 Η Η 0 H H < < 0 kJ < CO kJ 0 0 0 0 Ω CN Η ο Hi Μ Ω σι co ο <Ώ Ο Φ ο Μ 0 Ω Ω The 0 0 0 0 0 Η 0 H 0 0 O U CN 0 0 Hi Η 0 H 0 Ω 0 < 0 Ο 0 0 CO CO Η 0 Eh 0 Ç 0 0 0 Η 0 < d j — j Η CH Hi Μ Ω σ » σ> ο ω Ο ΓΟ CO Ω CN CN Μ ο Ω Ω The 0 Η 0 < Eh a Ω 1 0 0 0 Εη Η 0 ι 0 <c 0 0 E_l GGAAA 0 0 H Eh < 0 1 0 0 0 β-4 0 Ο Η 0 0 H 0 0 kJ 0 0 0 0 0 ΙΦ Ο φ 0 ΙΦ Ο · Φ σι CN 4J Pi Pi Ρ Q Ω φ β 0 0 ο 0 Ω gives 0 ΙΦ U > Η φ cn Η Φ Ρ Q Ω φ > Φ Φ φ > > Η f — ι Φ Φ [“*« · ο φ | —1 The na The β Ρ kO ko <Φ 4J φ CD Φ j— { β • Η •H & Φ φ φ Φ Φ φ Ο Ό Ό Ό co Φ Φ 0 0

338

Ω CO Η ο Hi Η Ω LO LT) j— | Ο Ui υ co Ω + co Μ ο Ω Ω ζ 0 Η H 0 H H < 0 0 0 < 0 < er ¹ H CO < Η 23 0 s H <3 0 0 CO H 0 Η 0 tr * rh 0 H 0 rd 0 0 κ Η 0 CJ rh H CO CJ rd H CO Q 0 0 0 (J r I 0 CO CO CJ Q_ | CO CO H U 0 Η H rh 0 H H rd < H 0 0 0 CJ < 0 CO CJ 0 0 < 0 0 0 0 H < H Ω co Η ο » ζ Μ Ω σ> 4 ¹ <£> Ο ω 0 O Ω co 'φ it's the Μ ο Ω Ω ζ 0 0 0 0 Eh 0 0 Η 0 H H 0 <J 0 0 0 0 H 0 H CO 0 < 0 H 0 (Ü Η 0 H CJ «Laugh * H CO U Ω 0 CO H H H 0 0 0 CO co Η 0 H 0 0 < 0 U 0 CO 0 0 < 0 0 Ω Η ο Hi Μ Ω CD CD CD Ο ω 0 CO co σ> Ω CO <N Η ο Ω Ω ζ 0 H 0 < < 0 0 0 r \ CO 0 H <τ C0 CJ <T CO CO < CO Η iCT H H H H 0 tó 0 Η ass 0 H CO 0 CJ CO 0 Q 0 < 0 0 Γ_ | 0 < H 0 0 Ο 0 0 Eh H · rh 0 0 CO H CO 0 0 CJ 0 < co H 0 0 0 1 <β υ- 0 iOS Ο · co Pi Q at CO 0 Ν 4J Pi 3 3 Q »» tn (β s 0 cx] Pi 0 at Ω 0 1-1 TO 0 0 THESE H σ> + σι H CD fal <+ co at Q Q Q > > at (D Φ TO at > > at Η ρ- | 0) [* ···> 0) [* «· ω P*·" υ at Γ — 1 X f— | ru (D ϋ Í4 <0 <0 U CD "1) 4-> at rd at j— | i "l ; 3 ΰ • Η •H at & 0) <D 0) <u CD •H <D ω υ ΤΟ TO TO TO <D TO ω at at TO 0 0 at 0

339

d co Η Οι ft ω Ω J — 1 j— | r ~ 1 ο <η 0 0 0 0 ο 0 0 0 W Ç ο Ω Ω ζ 0 < 0 0 0 0 0 CO 0 0 H H 0 < Εη Η CO Η r) Eh CD < 0 0 H Ε_ι CD co Eh 0 CD H Eh CO 0 Εη 0 0 Η 0 0 0 v_> Eh 0 < 0 kJ Eh co CO kJ Eh CD <C Μ 0 go ¹ 0 0 H 0 t-f * 0 co H CO 0 CD Eh kJ 0 FT < H 0 0 sL H 0 0 H 0 kJ 0 0 kJ 0 0 kJ 0 0 0 Eh < Eh 0 Eh Eh 0 Eh H Ω 0 Η α ft Μ Ω GRANDFATHER GRANDFATHER GRANDFATHER Ο ω 0 O O O Ω 0 0 0 Η Ç ο Ω Ω ζ 0 Eh < 0 0 Eh < 0 0 Eh 0 Η 0 Eh 0 Εη 0 Eh CO Εη 0 Eh CD 0 0 Eh 0 H 0 0 Eh 0 H 0 0 Eh 0 H 0 Eh Eh 0 Εη Eh < 0 Εη Eh < 0 Pi 0 ft 0 0 CD Eh co 0 CD Eh 0 0 Ω 0 Eh Eh H 0 Eh H Eh 0 Eh H Eh Ο < < < 0 0 0 0 0 0 0 CD 0 0 Eh 0 0 H < CO 0 Η CD 0 0 0 0 0 0 Ω ρ-j Η ο ft Μ Ω GRANDFATHER GRANDFATHER GRANDFATHER Ο w 0 σ> 0> ft Ω j— | J— { « Ο Ω Ω ζ 0 ' co CD 0 H co H 0 H 0 CO CD H H 0 H H 0 Eh H 0 Ρμ 0 0 0 0 0 0 0 0 0 Q 0 0 < CO CO 0 CD < Ο H 0 CO Η CD Eh H H 0 1 (0 ο (tí Ν 0 1 (0 O (0 4J & Z fc fc fc (0 g ϋ 0 Ω gives 0 1 (0 + + + θ ' Eh Εη Eh (0 CO fc CO O> fc CO laughs co 7 ¹ 00 GRANDFATHER 'φ co Εη M 0 1—1 | 0 l · —1 (you > M > > (you (Ç (0 75 75 75 (you (you (you (Ç Η ω Ρ ** ω [* ·. ω [** · υ (you φ Μ φ Μ φ α ft νο ft νο ft GRANDFATHER «Ρ 4J ,-1 Γ ~ Η rH The (ΰ <tí (you & The) • Η φ Ή φ •H Φ 0) O φ τ5 φ 75 Φ 75 ω τ5 75 75 φ (ΰ (you 0 0 0

340

ID No. SEQ. DA CDR3 221 446 221 CDR3 GACGGTATG GACGTC TCAGTAGTA ACTGGCGGC TACTACTAC GACGGTATG GACGTC ATCGTAGTA ACTGGCGGC TACTACTAC GACGGTATG GACGTC TCAGTAGTA ACTGGCGGC TACTACTAC ID No. SEQ. DA CDR2 206 206 206 CDR2 AGCACATCTC TGAAGAGC CACATTTTTT CGAATGACGA AAAATCCTAC AGCACATCTC TGAAGAGC H <O 0 Eh 0 <Eh CD Eh U Eh 0 0 Eh <0 Eh < Eh 0 0 <0 Eh Eh Eh U u 3 3 0 0 <0 3 0 Eh 0 O 3 < CACATTTTTT CGAATGACGA AAAATCCTAC ID No. SEQ. DA CDR1 196 196 196 CDR1 AATGCTAGA ATGGGTGTG AUG AATGCTAGA ATGGGTGTG AGC AATGCTAGA ATGGGTGTG AGC Mutation location FW FW, CR3 £ Variation E16Q + V24F + I83T + T119L ⁺ + + ⁺ ft O ft Eh 5 2 co 2 ft ft <M 00 2 Γ W> ^H ω I83K Center string 16H7 heavy chain 16H7 heavy chain 16H7 heavy chain

341

ID No. SEQ. DA CDR3 <0 221 GRANDFATHER 0 CJ 0 CD 0 0 CD CJ 0 H Eh CD Eh Eh CD H H CD Ç r_i 2 CD Eh C_i 0 CD Eh C_1 2 CD Eh Eh O 01 Eh CJ 0 Fh 0 0 Pm 0 kJ Eh CD CD kJ Eh CD CD kJ H CD W 0 0 CD H CD 0 CD H CD CD CD H 0 01 H 0 01 H 0 0 CJ H 0 kJ Eh 0 kJ 0 0 kJ H 0 0 <1 < H 01 Eh Eh 0 < H d The OJ H O Pi fa O GRANDFATHER <0 kO O U1 u O O O Q fa CN O1 O Q Q The 0 Eh < 0 0 Eh 0 0 Fh 0 Eh 0 Eh CD < H 0 Eh CD H 0 Eh CD 0 0 Eh 0 H 0 0 Rh 0 Fh 0 0 H 0 H cs Eh Eh < 0 H Eh < 0 H H 0 P4 <1 0 Eh CD 0 < 0 Eh CD 0 CD H CD 0 Q 0 Eh Eh Eh 0 Eh H H 0 Eh H H 0 < 0 0 0 CJ 0 0 CJ CD 0 0 Eh CD CD Eh CD 2 Eh CD < 0 0 0 0 0 0 d The H The < Pi M Ω GRANDFATHER GRANDFATHER GRANDFATHER O ω 0 σι ΟΊ σ Q w H H H O Q Q The CD CD CD 0 H 0 H 0 H CD < CD CD H H 0 H H 0 Fh H 0 CJ CD 0 CJ CD 0 CJ CD 0 M CD CD CD CD CD CD H CD H CD H CD Eh H Eh < 0 ird o · rd 0 tm The <d ΓΩ co 44 Pi g g Pi Q fa fa Q J 8 0 0 u 0 Ω gives 0 Md + d ~ + + + Ο » H <β Pi O Pi fa -rl 00 σ> σ 00 CTi O k 00 00 > 00 00 σ Φ> Q ft Q ft > rd rd rd 75 75 75 rd φ rd rd H | - ^ m ω P*". CO ["H O rd CD w 0) fa 0) fa k ft <0 ft GRANDFATHER ft GRANDFATHER <φ 44 f— | p-1 you rd rd rd & Φ •H OJ •H OJ Ή φ φ υ 0) 75 0) 75 (D 75 ω 75 75 75 rd rd rd 0 0 0

342

ID No. SEQ. DA CDR3 447 448 σι 0 O CD O O CD CD CD ft ft ft ft 0 ft CD < ft CD Ο 0 0 CD CD 0 CD ft you ft O ft CD CD CD CD CD U | ft CD 0 ft P5 0 LJ ft CD ft ft ft CD CD L> ft «D M 0 0 CD ft 0 0 CD ftl 0, «Id CD CD CD L-J HI <1 ft CD ft <1 ft CD CD FL 0 ft ft O O 0 CD 0 CD 0 L_J 0 01 ftl ft 0 ftl ft <1 U 0 Q . CN H ex Hi X Q LO LO σ> O X U O sji Q X Ç CN CN CN 0 Q Q The 0 ft CD CD ft O O CD O ft CD ft CD < ft CD ft CD ft CD CD ft O 0 ft O ft O CD ft CD ft CD CD <2 u CN ft ft < U ft ft CD ft CD Hi < 0 ft CD U CD ft CD CD < 0 ft CD O 0 ft ft ft U ft ft ft O CD U < < < 0 0 0 0 u CD CD u 0 CD ft CD 0 ft CD 0 ft CD CD ft < U O O O CD 0 < Q • J — I H Hi Hi H Q LO LO O O X U ΟΛ cn ’Φ Q H Ç H rd CN O Q Q The CD CD U < < 0 ft 0 ft CD CD CD CD <T. CD CD ft ft CD ft ft U ft ft tó O 0 0 CD CD 0 CD ft CD Q CD CD CD CD CD ft U ft CD ft CD CD O CD ft - = 5 ft CD CD 0 0 liti t> rd 0 Md CD (d CO ro ro 4J X X X 3 Q Q Q <0 g CD O U υ 0 x gives 0 o <d -H S100I election and Y107 D109S N92Q <0> Q CD CD Ό O OD rd CD id > rd ω ω OD LD O <d 0) X 0) X | - | The d The LO d LO CN "1) P p-1 j— | (d CN The rd CD •H & a> -t — 1 OD -H CD OD (ID <D u dD T5 0) T3 Ό nt X Ό Ό rd CD CD U U CD

343

ID No. SEQ. DA CDR3 450 451 452 CDR3 GTGGTA CAGGTGTGG GATAATACT GCTGATCAT GTGGTA ATATTATTA GTGGGAGCT TACTACTAC AGCGGTATG GACGTC TCAGTAGTA ACTGGCGGC TACTATTAC TACGACGGT ATGGACGTC ID No. SEQ. DA CDR2 249 206 206 CDR2 I GATGATAGCG ACCGGCCCTC THE Eh <U U Eh 0 <Eh 0 Eh 0 Eh 0 0 Eh <0 Η < Eh 0 U <0 Eh Eh Eh 0 U <<0 o <0 <0 Eh The 0 << CACATTTTTT CGAATGACGA AAAATCCTAC AGCACATCTC TGAAGAGC ID No. SEQ. DA CDR1 240 196 196 CDR1 0 GGGGGAAAC AACATTGGA AGGTGTGCA C AATGCTAGA ATGGGTGTG AGC AATGCTAGA ATGGGTGTG AGC ________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________1 Mutation location CDR3 CDR3 CDR3 Variation cn co C109S Y107 insertion Center string 22H5 light chain 22H5 heavy chain 16H7 heavy chain

344

Additionally, a hemibody was generated and studied. This structure was composed of the 16H7 light chain (L3; SEQ ID. NO: 50), which was paired with a genetically modified form of the 16H7 heavy chain; the genetically modified heavy chain was composed of the 16H7 heavy chain (SEQ ID. NO: 32) joined by means of a linker (G ₄ S) ₈ (SEQ ID. NO: 440) to a sequence of IgG2 Fc (SEQ ID. NO .: 441), which matched the 16H7 heavy chain Fc sequence. The component parts of the hemibody have the following sequences:

16H7 heavy chain:

MDMRVPAQLLGLLLLWLRGARCQVTLKESGPVLVKPTETLTLTCTVSGFSLNNARMGV SWIRQPPGKALEWLAHIFSNDEKSYSTSLKSRLTISKDTSKSQVVLIMTNMDPVDTATYY CARSWTGGYYYDGMDVWGQGTTVTVSSASTfCGPSVFPLAPCSRSTSESTAALGCLVK DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKP SNTKVDKTVERKSSVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRWSVLTWHQDWLNGKEYKCKVSNKG LPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGlSfVFSCSVMHEALHNHYTQKSLSLSP (SEQ ID NO:.. 32)

Linker:

GGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGGGGGGS (SEQ ID. N °: 440)

IgG2 Fc:

ERKSSVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYV DGVEVHNAKTKPREEQFNSTFRWSVLTVVHQDWLNGKEYKCKVSNKGLPAPTEKTIS KTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNVKTTPP MLDSDGSFFLYSKL.TVDKSRWQQGNVFSCSVMHEAI.HNHYTQKSLStSPGK (SEQ ID NO:.. 441)

The total heavy chain of the hemibody had the amino acid sequence shown below:

345 mdmrvpaqllgllllwlrgarcqvtlkesgpvlvkptetltltctvsgfslnnarmgv SW1RQPPGKALEWLAH1FSNDEKSYSTSLKSRLT1SKDTSKSQVVL1MTCMDPVDTATYY CARSVVTGGYYYDGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSÊSTAALGCLVK DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKP SNTKVDKTVERKSSVECPPCPAPPVAGPSVFLFPPKPKjDTLMISRTPEVTCVVVDVSHED PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVIYVVHQDWI.NGKEYKCKVSNKG LPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALEINHYTQKSLSLSP GGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSERKSSVECPPCPAPPV AGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTFRWSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPP SREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:.. 453), which is encoded by the following sequence:

ATGGACATGAGGGTGCCCGCTCAGCTCCTGGGGCTCCTGCTGCTGTGGCTGAGAGGT GCGCGCTGTCAGGTCACCTTGAAGGAGTCTGGTCCTGTGCTGGTGAAACCCACAGAG ACCCTCACGCTGACCTGCACCGTGTCTGGGTTCTCACTCAACAATGCTAGAATGGGT GTGAGCTGGATCCGTCAGCCCCCAGGGAAGGCCCTGGAGTGGCrTGCACACATTTTT TCGAATGACGAAAAATCCTACAGCACATCTCTGAAGAGCAGGCTCACCATCTCCAA ggacacctccaaaagccaggtggtcctaattatgaccaacatggaccctgtggaca cagcx'acatattactgtgcacggtcagtagtaactggc: ggctactactacgaCggta tggacgtctggggccaagggaccacggtcaccgtctctagtgccagcaccaagggc ccctccgtgttccctctggccccctgcagcagaagcaccagcgagagcacagccgcc CTGGGCTGCCTGGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCnTGGAACAGC ggagccctgaccagcggcgtgcacacctttccagccgtgctgcagagcagcggcct GTACAGCCTGAGCAGCGTGGTCACCGTGCCCAGCAGCAACTTCGGCACCCAGACCT acagctgtaacgtggaccacaagcccagcaacaccaaggtggacaagacagtgga GCGGAAGTCCAGCGTGGAGTGCCCTCCTTGTCCTGCCCCTCCTGTGGCCGGACCTAG cgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccgcga AGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGACCCCGAGGTGCAGTTCAATT ggtacgtggacggggtggaggtgcacaacgccaagaccaagccccgggaggaaca GTTCAACAGCACCTrCCGGGTG GTGTCCGTCCTCACCGTGGTGCACCAGGACTGGCT gaacggcaaagagtacaagtgcaaggtctccaacaagggcctgcctgcccccatcatgg AGAAAACCATCAGCAAGACCAAGGGCCAGCCTCGGGAGCCTCAGGTGTACÂCCCT

346

CCCCCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTCGTGAA GGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGA ACAACTACAAGACCACCCCCCCCATGCTGGACAGCGACGGCAGCTTCTTCCTGTACT CCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGTAGC GTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGTCTCCT ggcggaggcggaggatctggcggcggaggaagtggagggggcggatctggtggtg GAGGCAGCGGCGGAGGTGGAAGTGGCGGTGGAGGATCCGGTGGAGGCGGCTCAGG tggcggcggaagcgagagaaagtcctgcgtggagtgtccaccatgccctgctccac cagtggctggcccttccgtctttctctttccacctaaacctaaggatacactcatgat ctccagaactccagaggtcacatgtgtggtcgtcgatgtcagtcatgaggatcctga agtccagtttaactggtatgtggatggcgtcgaagtccataatgctaagacaaaacc tcgcgaagaacagtttaactccacctttagagtcgtgagcgtgctgacagtcgtcca tcaggattggctcaatgggaaagaatacaaatgtaaagtctctaacaaaggactgc ccgctcctatcgaaaagaccatctccaaaacaaaggggcagcccagagagccccag gtctacacactcccaccgtccagagaagagatgacaaaaaatcaggtgtcactcac ctgtctggtcaaggggttttacccctccgacattgccgtggaatgggaatccaatgg gcagcctgaaaacaattataagagtacacctcctatgctcgactctgatgggagttt CTTrCTCTACTCTAAACTCACAGT GGATAAGTCTAGATGGCAGCAGGGGAATGTCTT TTCCTGCTCCGTCATGCATGAAGCTCTCCACAATCATTATACACAGAAGTCTTTGTCC CTGTCCCCCGGCAAG (ID. SEQ. No.: 454)

Example 14.1

Β-Klotho binding ELISA for genetically modified antibodies

The genetically modified forms of 16H7 and 22H5 were tested for β-Klotho binding using an ELISA assay. The conditions for ELISA were as follows.

Maxisorp plates coated with streptavidin were incubated with 2 pg / ml β-Klotho overnight at 4 ° C. Antibodies were added in 3-fold serial dilutions starting at 1 μΜ for 1 hour at room temperature. Human HRP-conjugated anti-Fc was used as the detector antibody. The signal was developed with Lumiglo and read in

347

Envision.

The results of the ELISA assay are shown in Figure 32A-32C and indicate that most of the 16H7 variants have bound to human β-Klotho, except for a mutant that carries the tyrosine insert at position 107.

Example 14.2

Genetically modified variants of 16H7 and 22H5 bind to the native structure of the receptor, as demonstrated by FACS

A FACS binding assay was performed with several of the genetically modified forms of 16H7 and 22H5. The experiments were carried out as follows.

CHO cells that stably express the FGF21 receptor were treated with antibody-related 16H7 and 22H5 and also with genetically modified variants of them (1 pg per 1 x 10 ⁶ cells in 100 μΐ of PBS / 0.5% BSA). The cells were incubated with the antibodies at 4 ° C, followed by two washes with PBS / BSA. The cells were then treated with secondary antibodies labeled with FITC at 4 ° C, followed by two washes. The cells were resuspended in 1 ml of PBS / BSA and antibody binding was analyzed using a FACS Calibur instrument.

Consistent with the ELISA results, most of the genetically modified variants of FGF21 receptor agonist antibodies tested bind well to the cell surface FGF21 receptor in FACS. This observation further confirmed that the genetically guided modification of FGF21 receptor agonist antibodies maintains the link to the native structure. In a mutant, in which CDR3 was genetically modified to include tyrosine in the

At position Y107, a complete loss of binding to the receptor cell surface was observed, which is similar to the ELISA results. This observation highlights the role of the CDR3 loop in linking to native conformation.

Example 14.3

Activity of 16H7 and 22H5 variants in primary human adipocytes

FGF21 stimulates glucose uptake and lipolysis in cultured adipocytes and, therefore, adipocytes are often considered to be a physiologically relevant assay. A panel of the 16H7 and 22H5 genetically modified variants has been shown to exhibit FGF21-like Erk phosphorylation activity in the human adipocyte assay with an estimated EC50 of less than 15 than 10 nM, as shown in Table 7.

Table 7 - Variant activity in the human adipocyte assay

Center string ID No. IN SEQ. from jail variant Variant EC50 (nM) Heavy chain from 16H7 391 I83T 0.73 Heavy chain from 16H7 393 E16Q + V24F + I83T 0.38 Heavy chain from 16H7 398 D88R + P89A + V90E 0.35 Heavy chain from 16H7 394 E16Q + V24F + I83T + T119L 0.36 16H7 (WT) 0.53 Light chain 403 S94A 1.98

349

Center string ID No. IN SEQ. from jail variant Variant EC50 (nM) from 22H5 22H5 light chain 402 N92Q 3.33 Heavy chain from 16H7 400 Y107 deletion 1.04 Heavy chain from 16H7 396 I83K 0.39 Heavy chain from 16H7 397 S100I 0.17 Heavy chain from 16H7 401 D109S 0.31 Heavy chain from 16H7 399 D88R + P89A + V90E + S100I 0.14 Heavy chain from 16H7 395 E16Q + V24F + I83T + S100I + T119L 0.24 Heavy chain from 22H5 405 Y107 insertion 0.51 Heavy chain from 16H7 390 V24F 0.75 Heavy chain from 16H7 392 V24F + I83T 0.37 Light chain from 16H7 386 D49Y 0.60 16H7 light chain 387 D4 9A 0.63 16H7 light chain 389 D49A, D91A 1.4

350

Center string ID No. IN SEQ. from jail variant Variant EC50 (nM) 16H7 light chain 388 D91A 1.3 Light chain from 16H7 385 Q16K 0.11 22H5 (WT) 2.27

Example 14.4

Biacore connection experiments and off-rate measurement

The binding of 16H7 and 22H5 variants to human β-Klotho was tested using Biacore assays. Briefly, mouse anti-His antibody (Qiagen, Valencia, CA) was immobilized on a CM5 chip using amine coupling reagents (General Electronics, Piscataway, NJ). Recombinant human β-Klotho with His tag was captured in the second flow cell at approximately 100 RU. The first flow cell 10 was used as a background control. One hundred nM of mAbs were diluted in PBS plus 0.1 mg / ml BSA, P20 0.005%, and injected over the β-Klotho captured on the surface of the anti-His antibody. For kinetic measurement, 0.78-100 nM mAbs diluted in PBS plus 0.1 mg / ml BSA, 15 P20 0.005%, were injected on the β-Klotho surface.

The tested variants are summarized in Table 8:

351

Table 8 - Variants studied in connection and off-rate experiments.

SEQ ID NO. light chain CO H 14 14 H 403 402 H 14 SEQ ID NO. heavy chain CO 391 393 398 394 32 rd CO rd CO 400 σ CO 397 401 399 Light chain identifier / variation what ft L3 L3 L3 L3 L3 3 σ w ο CN CTl 3 L3 L3 L3 CO hl L3 Heavy chain identifier / variation ft I83T E16Q + V24F + I83T D88R + P89A + V90E E16Q + V24F + I83T + T119L H3 H2 H2 Y107 deletion I83K S100I D109S D88R + P89A + V90E + S100I Central antigen binding protein 16H7 16H7 16H7 16H7 16H7 22H5 22H5 16H7 16H7 16H7 16H7 16H7 Construction number 22H5 # 1, P60881.3 # 2, P60880.3 # 3, P60890.3 # 4, P60878.3 # 5, 16H7 WT # 6, P60898.3 # 7, P60897.3 # 8, P60886.3 m LD oo oo o <0 ft # 10, P60884.3 # 11, P60883.3 # 12, P60879.3

352

SEQ ID NO. light chain H H H kO 00 CO 387 389 00 00 CO 385 CO ID ID # SEQ. heavy chain 453 395 405 O ΟΪ CO 392 32 32 32 32 32 404 Light chain identifier / variation ! L3 CO i-q L3 CO h-q L3 (You Q Q ΟΊ Q + CTi Q σ Q ft CO O L2 Heavy chain identifier / variation Hemibody heavy chain E16Q + V24F + I83T + S100I + T119L Y107 insertion ft «tf 0 > V24F + I83T H3 H3 H3 H3 H3 C109S Central antigen binding protein 16H7 16H7 L 16H7 kO 16H7 16H7 16H7 16H7 16H7 16H7 22H5 Construction number # 13, P60882.3 # 14, P60891.3 # 15, P60889.3 CO 00 00 00 o k £) ass kO # 17, P60887.3 # 18, P60894.3 # 19, P60895.3 # 20, P60893.3 # 21, P60892.3 # 22, P60896.3 # 23, P60899.2

353

Among the genetically modified mAbs tested, most of them showed close connection to human β-Klotho, except # 15 which showed no link. Ά Table 9 below shows the binding of 100 nM mAbs to β-Klotho captured in anti-His. Figure 33 shows the comparison with off-rate.

Table 9 - Connection to β-Klotho

Sample koff (1 / s) # 20, P60893.3 1.9E-04 # 11, P60883.3 2.6E-04 # 23, P60899.2 3.0E-04 22H5 3.1E-04 # 18, P60894.3 3.1E-04 # 6, P60898.3 3.5E-04 # 13, P60882.3 4.4E-04 # 7, P60897.3 5.2E-04 # 8, P60886.3 5.3E-04

EXAMPLE 15

Combinations of antigen binding proteins show an additive effect

Antigen binding proteins that represent different binding compartments (bins) (Figures 11a and b) were selected and tested in pair-reporter assays to determine whether the pair of molecules would behave additively. The tests were carried out as follows.

On the first day, the AM-l / D FGFRlc + β-Klotho Luc clone was seeded in a 96-well plate at 20 K cells / well in DMEM + 10% FBS medium. The plate was incubated overnight. THE following day, the medium was replaced with medium

354 assay (DMEM + 0.2% FBS) and incubated overnight. From an antibody working stock (1 mg / ml in PBS), each antibody under study was prepared at a dilution of 2 pg / ml in assay medium. One hundred μΐ of every 5 antibodies to be tested were combined on a U-bottom plate. The assay medium was removed from the cells, and μΐ of the antibody mixtures were transferred to the cells. The antibody mixtures were incubated in the cells for 5 hours. Finally, each sample was read with Luciferase SteadyGlo reagent (50 μΐ / well), according to the manufacturer's specifications.

Table 10 below is a summary of the activity (% of FGF21 activity by the reporter-trial) observed by the study; Table 11 expresses the activities observed with respect to the compartments.

Table 10 - Antibody combination activity (%)

Iso 6 5 4 3 2 1 IgG2k 2G10 16H7 12E4.1 20D4.1 39F7 26H11.1 Iso IgG2k ND -1.1 23.5 25.4 12.5 9.2 17.9 1 26H11.1 17.9 19.1 36.7 21.4 28.3 20.7 2 39F7 9.2 9.1 37.0 30.8 21.4 3 20D4.1 12.5 13.5 19.4 32.0 4 12E4.1 25.4 28.8 41.5 5 16H7 23.5 27.8 6 2G10 -1.1

355

356

Surprisingly, several pairs of molecules showed an additive effect. As shown in Figures 34 and 35, respectively, 39F11 and FGF21 showed an additive effect when measured in the reporter test of Example 5, as did 16H7 and 39H11.

Summarizing the data from this set of experiments, it was observed that in antigen binding proteins from the same binding compartment, for example, 16H7 when paired with 20D4 (both from Group A), the added activity was not additive. This was also observed when 12E4 was paired with 26H11 (both from Group B). In addition, antigen-binding proteins paired from non-overlapping compartments showed additive activities, for example, 16H7 (Group A) paired with 26H11 or 12E4 (Group B), or paired with 39F7 (Group C). In addition, the 26H11 and 12E4 antigen binding proteins (Group B) showed an additive effect when combined with Group A, but not Group C, Abs, suggesting that there may be some overlap between Group B and Group C binding sites. and / or that the activation conformations induced by Group B and Group C antigen binding proteins are not mutually compatible. Finally, as expected, when a functional antigen-binding protein is paired with a non-functional antigen-binding protein (e.g., 2G10) that binds to a distinct, non-overlapping Group A, B, or C binding site, there is no effect on the activity of the Group A, B or C functional antigen binding protein.

Collectively, these data suggest that the revealed antigen binding proteins can be co-administered

357 to increase the effect that a certain antigen-binding protein can provide on its own.

Each reference cited here is incorporated by reference in its entirety in all its teachings 5 and for all purposes.

The present disclosure is not limited in scope by the specific modalities described here, which serve only as illustrations of individual aspects of the disclosure, and functionally equivalent methods and components form 10 aspects of the disclosure. In fact, several modifications of the disclosure, in addition to those shown and described here, will become evident to those skilled in the art from the description presented above and the accompanying drawings. These modifications are intended to be included within the scope of the appended claims.

Claims (5)

1. Isolated antibody, scFv or Fab, characterized by being able to bind to a complex comprising β-Klotho and FGFR1c and comprising the following CDRs: (a) CDRH1: SEQ ID NO: 122, CDRH2: SEQ ID NO: 133, CDRH3: SEQ ID NO: 148, CDRL1: NO: 176, and CDRL3: SEQ ID NO: (b) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 146, CDRL1: NO: 176, and CDRL3: SEQ ID NO: (c) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 147, CDRL1: NO: 176, and CDRL3: SEQ ID NO: (d) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 148, CDRL1: NO: 176, and CDRL3: SEQ ID NO: (e) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 150, CDRL1: NO: 174, and CDRL3: SEQ ID NO: (f) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 152, CDRL1: NO: 173, and CDRL3: SEQ ID NO: (g) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 152, CDRL1: NO: 173, and CDRL3: SEQ ID NO: (h) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 151, CDRL1: NO: 173, and CDRL3: SEQ ID NO: (i) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 153, CDRL1: SEQ ID NO: 166, CDRL2: SEQ ID 188; 122, CDRH2: SEQ ID NO: 133, SEQ ID NO: 167, CDRL2: SEQ ID 190; 122, CDRH2: SEQ ID NO: 133, SEQ ID NO: 167, CDRL2: SEQ ID 189; 122, CDRH2: SEQ ID NO: 134, SEQ ID NO: 166, CDRL2: SEQ ID 188; 124, CDRH2: SEQ ID NO: 136, SEQ ID NO: 161, CDRL2: SEQ ID 183; 124, CDRH2: SEQ ID NO: 138, SEQ ID NO: 162, CDRL2: SEQ ID 184; 124, CDRH2: SEQ ID NO: 139, SEQ ID NO: 162, CDRL2: SEQ ID 186; 124, CDRH2: SEQ ID NO: 137, SEQ ID NO: 164, CDRL2: SEQ ID 186; 131, CDRH2: SEQ ID NO: 140, SEQ ID NO: 163, CDRL2: SEQ ID Petition 870190097789, of 09/30/2019, p. 8/13 2/4 NO: 173, and CDRL3: SEQ ID NO: (j) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 149, CDRL1: NO: 172, and CDRL3: SEQ ID NO: (k) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 145, CDRL1: NO: 171, and CDRL3: SEQ ID NO: (l) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 154, CDRL1: NO: 171, and CDRL3: SEQ ID NO: (m) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 157, CDRL1: NO: 177, and CDRL3: SEQ ID NO: (n) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 156, CDRL1: NO: 173, and CDRL3: SEQ ID NO: (o) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 156, CDRL1: NO: 173, and CDRL3: SEQ ID NO: (p) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 155, CDRL1: NO: 178, and CDRL3: SEQ ID NO: (q) CDRH1: SEQ IDNO: CDRH3: SEQ ID NO: 156, CDRL1: NO: 179, and CDRL3: SEQ ID NO: 2. Isolated antibody, being able to bind to a complex that comprises 185; 123, CDRH2: SEQ ID NO: 135, SEQ ID NO: 159, CDRL2: SEQ ID 181; 121, CDRH2: SEQ ID NO: 132, SEQ ID NO: 158, CDRL2: SEQ ID 180; 126, CDRH2: SEQ ID NO: 133, SEQ ID NO: 168, CDRL2: SEQ ID 191; 130, CDRH2: SEQ ID NO: 144, SEQ ID NO: 169, CDRL2: SEQ ID 192; 129, CDRH2: SEQ ID NO: 142, SEQ ID NO: 163, CDRL2: SEQ ID 194; 128, CDRH2: SEQ ID NO: 141, SEQ ID NO: 163, CDRL2: SEQ ID 194; 127, CDRH2: SEQ ID NO: 135, SEQ ID NO: 170, CDRL2: SEQ ID 193; or 128, CDRH2: SEQ ID NO: 143, SEQ ID NO: 163, CDRL2: SEQ ID 194. scFv or Fab, characterized by β-Klotho and FGFR1c, wherein said isolated antibody, scFv or Fab comprises (a) SEQ ID NO: 50 and SEQ ID NO: 68, Petition 870190097789, of 09/30/2019, p. 9/13 3/4 (B) SEQ ID AT THE: 48 and SEQ ID AT THE: 66, (ç) SEQ ID AT THE: 49 and SEQ ID AT THE: 67, (d) SEQ ID AT THE: 50 and SEQ ID AT THE: 69, (and) SEQ ID AT THE: 51 and SEQ ID AT THE: 70, 5 (f) SEQ ID AT THE: 52 and SEQ ID AT THE: 71, (g) SEQ ID AT THE: 53 and SEQ ID AT THE: 72, (H) SEQ ID AT THE: 54 and SEQ ID AT THE: 73, (i) SEQ ID AT THE: 56 and SEQ ID AT THE: 74, (j) SEQ ID AT THE: 57 and SEQ ID AT THE: 76, 10 (k) SEQ ID AT THE: 59 and SEQ ID AT THE: 78, (l) SEQ ID AT THE: 60 and SEQ ID AT THE: 79, (m) SEQ ID AT THE: 61 and SEQ ID AT THE: 80, (n) SEQ ID AT THE: 62 and SEQ ID AT THE: 81, (O) SEQ ID AT THE: 63 and SEQ ID AT THE: 82, 15 (P) SEQ ID AT THE: 64 and SEQ ID AT THE: 83, or (q) SEQ ID AT THE: 65 and SEQ ID AT THE: 84. 3. Isolated antibody, scFv or Fab, characterized by being able to bind to a complex comprising β-Klotho and FGFR1c, in which said antibody, scFv or Fab comprises 20 (The) SEQ ID AT THE: 14 and SEQ ID AT THE: 32 (B) SEQ ID AT THE: 12 and SEQ ID AT THE: 30 (ç) SEQ ID AT THE: 13 and SEQ ID AT THE: 31 (d) SEQ ID AT THE: 14 and SEQ ID AT THE: 33 (and) SEQ ID AT THE: 15 and SEQ ID AT THE: 34 25 (f) SEQ ID AT THE: 16 and SEQ ID AT THE: 35 (g) SEQ ID AT THE: 17 and SEQ ID AT THE: 36 (H) SEQ ID AT THE: 18 and SEQ ID AT THE: 37 (i) SEQ ID AT THE: 20 and SEQ ID AT THE: 38 (j) SEQ ID AT THE: 21 and SEQ ID AT THE: 40 30 (k) SEQ ID AT THE: 23 and SEQ ID AT THE: 41 Petition 870190097789, of 09/30/2019, p. 10/13 4/4 (l) SEQ ID AT THE: 24 and SEQ ID AT THE: 42, (m) SEQ ID AT THE: 25 and SEQ ID AT THE: 46, (n) SEQ ID AT THE: 26 and SEQ ID AT THE: 47, (O) SEQ ID AT THE: 27 and SEQ ID AT THE: 44, P) SEQ ID AT THE: 28 and SEQ ID AT THE: 43, or (q) SEQ ID AT THE: 29 and SEQ ID AT THE: 45. An isolated antibody, scFv or Fab, according to claim 1, characterized in that the antigen binding protein is a human antibody, a humanized antibody, a chimeric antibody, a monoclonal antibody, a recombinant antibody, a fragment of binding antibody antigen, a single chain antibody, a diabody, a triabody, a tetrabody, a Fab fragment, an F (fab ') 2 fragment, a domain antibody, an IgD antibody, an IgE antibody, an IgM antibody, an antibody IgG1, an IgG2 antibody, an IgG3 antibody, an IgG4 antibody or an IgG4 antibody that has at least one mutation in the hinge region. Pharmaceutical composition, characterized in that it comprises the isolated antibody, scFv or Fab, as defined in any one of claims 1 to 4, in admixture with a pharmaceutically acceptable carrier.